9-(piperazinylalkyl) carbazoles as bax-modulators

ABSTRACT

The present invention is related to piperazine derivatives of carbazole of formula (I) notably for use as pharmaceutically active compounds, as well as to pharmaceutical formulations containing such piperazine derivatives of carbazole. Said piperazine derivatives of carbazole are efficient modulators, in particular efficient inhibitors, of the Bax function and/or activation. The present invention is furthermore related to novel piperazine derivatives of carbazole as well as methods of their preparation.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a 371 of PCT/IB00/01497 filed Oct. 18, 2000.

FIELD OF THE INVENTION

The present invention is related to piperazine derivatives of carbazolenotably for use as pharmaceutically active compounds, as well aspharmaceutical formulations containing such piperazine derivatives ofcarbazole useful for the treatment of disorders associated withapoptosis, including neurodegenerative disorders, diseases associatedwith polyglutamine tracts, epilepsy, ischemia, infertility,cardiovascular disorders, renal hypoxia, hepatitis and AIDS. Saidpiperazine derivatives of carbazole display a modulatory and mostnotably an inhibitory activity of the cellular death agonist Bax and theactivation pathways leading to Bax and allows therefore to block therelease of cytochrome c. The present invention is furthermore related tonovel piperazine derivatives of carbazole as well as to methods of theirpreparation.

BACKGROUND OF THE INVENTION

Apoptosis denotes the complex contortions of the membrane and organellesof a cell as it undergoes the process of programmed cell death. Duringsaid process, the cell activates an intrinsic suicide program andsystematically destroys itself in a controlled manner or by aself-regulated process. The following series of events can be observed:

-   -   The cell surface begins to bleb and expresses pro-phagocytic        signals. The whole apoptotic cell then fragments into        membrane-bound vesicles that are rapidly and neatly disposed of        by phagocytosis, so that there is minimal damage to the        surrounding tissue.    -   The cell then separates from its neighbors.    -   The nucleus also goes through a characteristic pattern of        morphological changes as it commits genetic suicide. The        chromatin condenses and is specifically cleaved to fragments of        DNA.

Neuronal cell death plays an important role in ensuring that the nervoussystem develops normally. It appears that the death of developingneurons depends on the size of the target that they innervate: cellswith fewer synaptic partners are more likely to die than those that haveformed multiple synapses. This may reflect a process, which balances therelative number of pre- to postsynaptic neurons in the developingnervous system. Although neuronal cell death is assumed to be apoptotic,it is only recently that neurons in developing rodent brain wereconclusively shown to undergo apoptosis as classified by morphology andDNA fragmentation.

Neuronal death occurs via either apoptotic or necrotic processesfollowing traumatic nerve injury or during neurodegenerative diseases.Multiple components are emerging as key players having a role in drivingneuronal programmed cell death. Amongst the components leading toneuronal apoptosis are protein members belonging to the Bcl-2 family(see Jacobson, M. D. 1997. Current Biology 7:R 277-R281; Kroemer, G. C.1997. Nature Medicine: 614-620; Reed, J. C. 1997. Nature 387:773-776).

Bcl-2 is a 26 kDa protein that localizes to the mitochondrial,endoplasmatic reticulum and perinuclear membranes. It is known by aperson skilled in the art that the entire Bcl-2 family comprises bothanti-apoptotic (Bcl-2, Bcl-x_(L), Bcl-w, Mcl-1, A1, NR-13, BHRF1,LMW5-HL, ORF16, KS-Bcl-2, E1B-19K, CED-9) and pro-apoptotic (Bax, Bak,Bok, Bik, Blk, Hrk, BNIP3, Bim_(L), Bad, Bid, EGL-1) molecules (seeKelekar, A., and C. B. Thompson 1998. Trends in Cell Biology 8:324-330).Said proteins can form homo- and hetero-dimers that involve amino acidsequences known as Bcl-2 homology (BH) domains. So far, four of saiddomains (BH1 to 4) have been identified, the BH3 having been attributeda particularly prominent role in view of the death-promoting cascade.Said BH3 domain of the pro-apoptotic members appears to be required forthe interaction between anti and pro-apoptotic molecules. The principalsite of action of some of the Bcl-2 family members seems to be themitochondria. Mitochondria have been shown to play a major role in manytypes of apoptosis. In particular, this organelle has been shown torelease Apoptosis Inducing Factor and cytochrome c, a hemoprotein whichis bound to the outer surface of the inner mitochondrial membrane. Saidcytochrome c has been shown to trigger caspase 9 activation throughApaf-1/caspase 9 complex formation. Bcl-2 family members play a key rolein regulating cytochrome c release. While Bcl-2 and Bcl-x_(L) have beenshown to suppress cytochrome c release, Bax has been found to stimulatethis event both in vitro using isolated mitochondria as well as inintact cells following heterologous expression (Martinou et al.; 1995The Journal of Cell Biology, 128, 201-208). The mechanisms by whichthese proteins perform their function are currently unknown. Thethree-dimensional structure of Bcl-xL and Bid revealed structuralsimilarities between these proteins and the channel-forming domains ofthe bacterial toxins colicins and diphtheria toxins. Consistent withsuch structural similarity, some members of this family including Baxwere also found able to form ion channels in synthetic lipid membranes.The channel forming activity of these proteins has not yet beendemonstrated in vivo.

Studies performed with Bax-deficient mice led to the conclusion that Baxplays a prominent role within the apoptosis pathways, notably inneuronal apoptosis. Bax is viewed to be essential for apoptosis inducedby NGF deprivation in neonatal sympathetic neurons or for apoptosisinduced in cerebellar granule cells by potassium deprivation from theculture medium. Moreover, it was found that in the Bax-deficient mice(knock-out) neonatal moto-neurons from the facial nucleus can survivefollowing axotomy (see Deckwerth, T. L., Elliott J. L., Knudson C. M. etal. 1996. Neuron 17, 401-41). Hence, the inhibition of the Bax activityleading to the prevention of cytochrome c release from mitochondriaduring apoptosis, is viewed to be useful to protect neurons and alsoother cell types from various cell death stimuli.

In WO 97/01635 (Neurex Corp.) the inhibition of apoptosis in an effortto promote cell survival is suggested to be achieved by introducing intothe cell a chimeric gene containing a polynucleotide encoding aBax-ω-polypeptide being operably linked to a promoter effective to causetranscription of the polynucleotide in the cell. It is reported that theexpression of the Bax-ω-polypeptide is effective to inhibit apoptosis inthe cell.

WO 96/06863 claims agents for inducing apoptosis, notably for cancertherapy. Such agents interact with extracellular or cell surfacemembrane bound opiod-like molecules or their receptors. Such agents maybe coupled to peptides which assist in the transport of the agentsthrough the cell membrane to promote internalisation and accumulation inthe cell nucleus if this is the site at which the agent producesapoptosis.

Perez et al. in Nat. Genet. 1999, 21(2), 200-203 have indicated thatapoptosis plays a fundamental role in follicular atresia and theysuggest to selectively disrupt the Bax function in order to extend theovarian lifespan.

Bax inhibition could indeed represent an interesting therapy for alldiseases associated with apoptosis, including neurodegenerative diseases(e.g. Alzheimer's disease, Parkinson's disease, diseases associated withpolyglutamine tracts including Huntington's disease, spinocerebellarataxias and dentatorubral-pallidoluysian atrophy; amyotrophic lateralsclerosis, retinitis pigmentosa and multiple sclerosis, epilepsy),ischemia (stroke, myocardial infarction and reperfusion injury),infertility (like pre-mature menopause, ovarian failure or follicularatresia), cardiovascular disorders (arteriosclerosis, heart failure andheart transplantation), renal hypoxia, hepatitis and AIDS.

Hence, it is an objective of the present invention to provide compoundsenabling the treatment of a whole variety of apoptosis-relateddisorders, including notably the above mentioned diseases.

It is specifically an objective of the present invention to provide atreatment of apoptosis related disorders by specifically modulating,e.g. by inhibiting, the Bax function or by inhibiting the Baxactivation.

It is notably an objective of the present invention to providerelatively small molecule pharmaceuticals, more specificallynon-proteinaceous molecules that avoid essentially all of the drawbacksarising from the use of large bio-peptides or bio-proteins (e.g. theirrestricted bio-availability as well as problems arising from possible invivo intolerance), however, which are suitable for the treatment ofdiseases associated with abnormal apoptosis. It is particularly anobjective of the present invention to provide relatively small moleculechemical compounds which are suitable Bax modulators (e.g. compoundsinhibiting the Bax function or inhibiting the Bax activation) so to beavailable for a convenient method of treating diseases involvingabnormal apoptosis. Moreover, it is an objective of the presentinvention to provide methods for preparing said small molecule chemicalcompounds. It is furthermore an objective of the present invention toprovide new pharmaceutical formulations for the treatment of diseaseswhich are caused by abnormal apoptosis, more specifically by Bax. It isfinally an objective of the present invention to provide a method oftreating diseases that are caused by abnormal apoptosis.

DESCRIPTION OF THE INVENTION

The aforementioned objectives have been met according to the independentclaims which are set out hereinafter in the description. Preferredembodiments are set out within the dependent claims.

The following paragraphs provide definitions of the various chemicalmoieties that make up the compounds according to the invention and areintended to apply uniformly throughout the specification and claimsunless an otherwise expressly set out definition provides a broaderdefinition.

“C₁-C₆-alkyl” refers to monovalent alkyl groups having 1 to 6 carbonatoms. This term is exemplified by groups such as methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-hexyl and thelike.

“Aryl” refers to an unsaturated aromatic carbocyclic group of from 6 to14 carbon atoms having a single ring (e.g. phenyl) or multiple condensedrings (e.g. naphthyl). Preferred aryl include phenyl, naphthyl,phenantrenyl and the like.

“C₁-C₆-alkyl aryl” refers to C₁-C₆-alkyl groups having an arylsubstituent, including benzyl, phenethyl and the like.

“Heteroaryl” refers to a monocyclic heteroaromatic, or a bicyclic or atricyclic fused-ring heteroaromatic group. Particular examples ofheteroaromatic groups include optionally substituted pyridyl, pyrrolyl,furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl,isothiazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl,1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl,1,3,4-oxadiazolyl, 1,3,4-triazinyl, 1,2,3-triazinyl, benzofuryl,[2,3-dihydro]benzofuryl, isobenzofuryl, benzothienyl, benzotriazolyl,isobenzothienyl, indolyl, isoindolyl, 3H-indolyl, benzimidazolyl,imidazo[1,2-a]pyridyl, benzothiazolyl, benzoxazolyl, quinolizinyl,quinazolinyl, pthalazinyl, quinoxalinyl, cinnnolinyl, napthyridinyl,pyrido[3,4-b]pyridyl, pyrido[3,2-b]pyridyl, pyrido[4,3-b]pyridyl,quinolyl, isoquinolyl, tetrazolyl, 5,6,7,8-tetrahydroquinolyl,5,6,7,8-tetra-hydroisoquinolyl, purinyl, pteridinyl, carbazolyl,xanthenyl or benzoquinolyl.

“C₁-C₆-alkyl heteroaryl” refers to C₁-C₆-alkyl groups having aheteroaryl substituent, including 2-furylmethyl, 2-thienylmethyl,2-(1H-indol-3-yl)ethyl and the like.

“Alkenyl” refers to alkenyl groups preferably having from 2 to 6 carbonatoms and having at least 1 or 2 sites of alkenyl unsaturation.Preferable alkenyl groups include ethenyl (—CH═CH₂), n-2-propenyl(allyl, —CH₂CH═CH₂) and the like.

“Alkynyl” refers to alkynyl groups preferably having from 2 to 6 carbonatoms and having at least 1-2 sites of alkynyl unsaturation, preferredalkynyl groups include ethynyl (—C≡CH), propargyl (—CH₂C≡CH), and thelike.

“Acyl” refers to the group —C(O)R where R includes “C₁-C₆-alkyl”,“aryl”, “heteroaryl”, “C₁-C₆-alkyl aryl” or “C₁-C₆-alkyl heteroaryl”.

“Acyloxy” refers to the group —OC(O)R where R includes “C₁-C₆-alkyl”,“aryl”, “hetero-aryl”, “C₁-C₆-alkyl aryl” or “C₁-C₆-alkyl heteroaryl”.

“Alkoxy” refers to the group —O—R where R includes “C₁-C₆-alkyl” or“aryl” or “hetero-aryl” or “C₁-C₆-alkyl aryl” or “C₁-C₆-alkylheteroaryl”. Preferred alkoxy groups include by way of example, methoxy,ethoxy, propoxy, butoxy, phenoxy and the like.

“Alkoxycarbonyl” refers to the group —C(O)OR where R includes“C₁-C₆-alkyl” or “aryl” or “heteroaryl” or “C₁-C₆-alkyl aryl” or“C₁-C₆-alkyl heteroaryl”.

“Aminocarbonyl” refers to the group —C(O)NRR′ where each R, R′ includesindependently hydrogen or C₁-C₆-alkyl or aryl or heteroaryl or“C₁-C₆-alkyl aryl” or “C₁-C₆-alkyl heteroaryl”.

“Acylamino” refers to the group —NR(CO)R′ where each R, R′ isindependently hydrogen or “C₁-C₆-alkyl” or “aryl” or “heteroaryl” or“C₁-C₆-alkyl aryl” or “C₁-C₆-alkyl heteroaryl”.

“Halogen” refers to fluoro, chloro, bromo and iodo atoms.

“Sulfonyl” refers to group “—SO₂—R” wherein R is selected from H,“aryl”, “heteroaryl”, “C₁-C₆-alkyl”, “C₁-C₆-alkyl” substituted withhalogens e.g. an —SO₂—CF₃ group, “C₁-C₆-alkyl aryl” or “C₁-C₆-alkylheteroaryl”.

“Sulfoxy” refers to a group “—S(O)—R” wherein R is selected from H,“C₁-C₆-alkyl”, “C₁-C₆-alkyl” substituted with halogens e.g. an —SO—CF₃group, “aryl”, “heteroaryl”, “C₁-C₆-alkyl aryl” or “C₁-C₆-alkylheteroaryl”.

“Thioalkoxy” refers to groups —S—R where R includes “C₁-C₆-alkyl” or“aryl” or “hetero-aryl” or “C₁-C₆-alkyl aryl” or “C₁-C₆-alkylheteroaryl”. Preferred thioalkoxy groups include thiomethoxy,thioethoxy, and the like.

“Substituted or unsubstituted”: Unless otherwise constrained by thedefinition of the individual substituent, the above set out groups, like“alkyl”, “alkenyl”, “alkynyl”, “aryl” and “heteroaryl” etc. groups mayoptionally be substituted with from 1 to 5 substituents selected fromthe group consisting of “C₁-C₆-alkyl”, “C₁-C₆-alkyl aryl”, “C₁-C₆-alkylheteroaryl”, “C₂-C₆-alkenyl”, “C₂-C₆-alkynyl”, primary, secondary ortertiary amino groups or quarter-nary ammonium moieties, “acyl”,“acyloxy”, “acylamino”, “aminocarbonyl”, “alkoxycarbonyl”, “aryl”,“heteroaryl”, carboxyl, cyano, halogen, hydroxy, mercapto, nitro,sulfoxy, sulfonyl, alkoxy, thioalkoxy, trihalomethyl and the like.Alternatively, said substitution could also comprise situations whereneighboring substituents have undergone ring closure, notably whenviccinal functional substituents are involved, thus forming e.g.lactams, lactons, cyclic anhydrides, but also acetals, thioacetals,aminals formed by ring closure for instance in an effort to obtain aprotective group.

“Pharmaceutically acceptable salts or complexes” refers to salts orcomplexes of the below-identified compounds of formula I that retain thedesired biological activity. Examples of such salts include, but are notrestricted to acid addition salts formed with inorganic acids (e.g.hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid,nitric acid, and the like), and salts formed with organic acids such asacetic acid, trifluoroacetic acid, oxalic acid, tartaric acid, succinicacid, malic acid, fumaric acid, maleic acid, ascorbic acid, benzoicacid, tannic acid, pamoic acid, alginic acid, polyglutamic acid,naphthalene sulfonic acid, naphthalene disulfonic acid, andpolygalacturonic acid. Said compounds can also be administered aspharmaceutically acceptable quaternary salts known by a person skilledin the art, which specifically include the quarternary ammonium salt ofthe formula —NR,R′,R″⁺Z⁻, wherein R, R′, R″ is independently hydrogen,alkyl, or benzyl, and Z is a counterion, including chloride, bromide,iodide, —O-alkyl, toluenesulfonate, methylsulfonate, sulfonate,phosphate, or carboxylate (such as benzoate, succinate, acetate,glycolate, maleate, malate, fumarate, citrate, tartrate, ascorbate,cinnamoate, mandeloate, and diphenylacetate).

“Pharmaceutically active derivative” refers to any compound that uponadministration to the recipient, is capable of providing directly orindirectly, the activity disclosed herein.

“Enantiomeric excess” (ee) refers to the products that are obtained byan essentially enantiomeric synthesis or a synthesis comprising anenantioselective step, whereby a surplus of one enantiomer in the orderof at least about 52% ee is yielded. In the absence of an enantiomericsynthesis, racemic products are usually obtained that do however alsohave the inventive set out activity as Bax inhibitors.

Quite surprisingly, it was now found that the piperazine derivatives ofcarbazole according to formula I are suitable pharmaceutically activeagents, notably by effectively modulating the Bax function or the Baxactivation.

Compounds of formula I according to the present invention are thosewherein

R⁰ and R¹ are selected independently from each other from substituentsincluding or consisting of hydrogen, halogen, cyano, sulfonyl, sulfoxy,substituted or unsubstituted C₁-C₆-thioalkoxy, nitro, primary, secondaryor tertiary amine or sulfonamide, aminocarbonyl, aminothiocarbonyl,hydroxy, substituted or unsubstituted C₁-C₆-alkoxy, aryloxy,heteroaryloxy, carboxylic amide, alkoxycarbonyl, carboxylic ester,carboxylic acid, substituted or unsubstituted C₁-C₆-alkyl carbonyl,substituted or unsubstituted arylcarbonyl or hetero-arylcarbonyl,substituted or unsubstituted saturated or unsaturatedC₃-C₈-cycloalkylcarbonyl, substituted or unsubstituted C₁-C₆-alkyl,substituted or unsubstituted C₂-C₆-alkenyl, substituted or unsubstitutedC₂-C₆-alkynyl, substituted or unsubstituted aryl or heteroaryl,substituted or unsubstituted 3-8 membered saturated or unsaturatedcyclic alkyl.

R² is selected from the group comprising or consisting of hydrogen,substituted or unsubstituted C₁-C₆-alkyl, substituted or unsubstitutedC₂-C₆-alkenyl, substituted or unsubstituted C₂-C₆-alkynyl, substitutedor unsubstituted aryl or heteroaryl, substituted or unsubstituted 3-8membered saturated and unsaturated cyclic alkyl, sulfoxy, sulfonyl,sulfonamide, carboxylic amide, aminocarbonyl, alkoxycarbonyl, hydrazineacyl, substituted or unsubstituted carbonyl-C₁-C₆-alkyl, substituted orunsubstituted arylcarbonyl or heteroarylcarbonyl, substituted orunsubstituted saturated or unsaturated C₃-C₈-cycloalkylcarbonyl, alkoxy,C₁-C₆-thioalkoxy.

R³ is selected from the group comprising or consisting of hydrogen,halogen, substituted or unsubstituted C₁-C₆-alkyl, substituted orunsubstituted C₂-C₆-alkenyl, substituted or unsubstituted C₂-C₆-alkynyl,substituted or unsubstituted aryl or heteroaryl, substituted orunsubstituted 3-8 membered saturated and unsaturated cyclic alkyl,alkoxycarbonyl, carboxylic amide, C₁-C₆-alkoxy, substituted orunsubstituted aryloxy, substituted or unsubstituted hetero-aryloxy,hydroxy, substituted or unsubstituted C₁-C₆-alkyl carbonyl, substitutedor unsubstituted arylcarbonyl or heteroarylcarbonyl, substituted orunsubstituted saturated or unsaturated C₄-C_(g)-cycloalkylcarbonyl, orR³ could be an oxo (═O) group.

k and l are independently from each other an integer from 0 to 4,preferably they are both independently from each other between 0 and 2and most preferred either 0 or 1.

X is a substituted methylene group, i.e. a group of the formula—(CR′R″)—, whereby at least one of R′ and/or R″ is not hydrogen but asubstituent containing at least one heteroatom. Thus, R′ and/or R″ isselected from the group comprising or consisting of a substitutedC₁-C₆-alkyl, an unsubstituted or substituted aryl or heteroaryl, asubstituted or unsubstituted C₁-C₆-alkoxy, substituted or unsubstitutedC₂-C₆-alkenyl, substituted or unsubstituted C₂-C₆-alkynyl, a primary,secondary or tertiary amino, a quarternary ammonium salt of the formula—NR,R′,R″⁺Z⁻, wherein R, R′, R″ is independently hydrogen, alkyl, orbenzyl, and Z is a counterion, including chloride, bromide, iodide,—O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, orcarboxylate (such as benzoate, succinate, acetate, glycolate, maleate,malate, fumarate, citrate, tartrate, ascorbate, cinnamoate, mandeloate,and diphenylacetate); or R′ and/or R″ could be an acylamino,amino-carbonyl, C₁-C₆-alkoxycarbonyl, a carboxylic ester acid or amide,halogen, hydroxy, sulfonyl, sulfonamide, C₁-C₆-thioalkyl,C₁-C₆-thioalkoxy or X could be a C═O or a C═S group.

m and n are independently from each other an integer from 1 to 3,preferably both m and n are 1. o is an integer from 0 to 8. Preferably ois an integer from 0 to 2, most preferred is o=0 or 2.

Specifically, R⁰ and R¹ of formula I are selected independently fromeach other of the group comprising or consisting of hydrogen, halogen,cyano, substituted or unsubstituted and C₁-C₆-alkyl, substituted orunsubstituted 3-8 membered saturated or unsaturated cyclic alkyl,C(═O)ORa, C(═O)NRaRb, C(═O)NRaRc, C(═O)Ra, C(═O)Rc, CRa(═N—N-Rb),CRa(═N—N-Rc), CRa(═N—O-Rb), C₁-C₆-alkylene, trifluoromethyl,trifluoromethoxy, ORa, ORc, NRaRb, NRaRc, NRaC(═O)NRaRb, NRaC(═O)NRaRc,NRaC(═O)Rb, SRa, SRc, NRaC(═O)Rc, OC(═O)Ra, OC(═O)Rc, NRa(SO₂Rb),NRa(SO₂Rc), SO₂NRaRb, SO₂NRaRc, NO₂, CH₂NRaRb, CH₂NRaRc,CH₂NRaC(═O)NRaRb, CH₂NRaC(═O)NRaRc, CH₂NRaC(═O)Rb, CH₂NRaC(═O)Rc,CH₂NRa(SO₂Rb), CH₂NRa(SO₂Rc), OSO₂-trifluoromethyl.

Alternatively, R⁰ and R¹ according to formula I could also beindependently from each other selected from an aryl or a 5-6-memberedheterocyclic group containing at least one hetero atom selected fromoxygen, nitrogen and sulfur, being optionally substituted by at leastone C₁-C₆-alkyl, C(═O)ORa, C₁-C₆-alkylene, trifluoromethyl,trifluoromethoxy, ORa, OC(═O)Ra, OC(═O)Rc, NRaRb, CH₂—NRaRb, SRa, SRc,NO₂, cyano, halogen, SO₂NRaRb, SO₂NRaRc, NRaSO₂Ra, NRaSO₂Rc,C₁-C₄-alkylene C(═O)ORa, OSO₂-trifluoromethyl.

According to a further preferred embodiment R² within formula I isselected from the group comprising or consisting of hydrogen,substituted or unsubstituted C₁-C₆-alkyl, substituted or unsubstituted3-8 membered saturated cyclic alkyl, C(═O)ORa, C(═O)NRaRb, C(═O)NRaRc,C(═O)Ra, C(═O)Rc, CRa(═N—N-Rb), CRa(═N—N-Rc), CRa(═N—O-Rb),C₁-C₆-alkylene, trifluoromethyl, trifluoromethoxy, RaC(═O)NRaRc,RaC(═O)Rb, RaC(═O)Rc, Ra(SO₂Rb), Ra(SO₂Rc), SO₂NRaRb, SO₂NRaRc,CH₂NRaRb, CH₂NRaRc, CH₂NRaC(═O)NRaRb, CH₂NRaC(═O)NRaRc, CH₂NRaC(═O)Rb,CH₂NRaC(═O)Rc, CH₂NRa(SO₂Rb), CH₂NRa(SO₂Rc), OSO₂-trifluoromethyl.

Alternatively, R² within formula I could also be independently from eachother selected from C₁-C₆-alkylaryl, C₁-C₆-alkylheteroaryl, an aryl or a5-6-membered heterocyclic group containing at least one hetero atomselected from oxygen, nitrogen and sulfur, both the aryl, heterocyclicgroup being optionally substituted by at least one C₁-C₆-alkyl,C(═O)ORa, C₁-C₆-alkylene, trifluoromethyl, trifluoromethoxy, ORa,OC(═O)Ra, OC(═O)Rc, NRaRb, CH₂—NRaRb, NO₂, cyano, halogen, SO₂NRaRb,SO₂NRaRc, NRaSO₂Ra, NRaSO₂Rc, C₁-C₆-allyleneC(═O)ORa,OSO₂-trifluoromethyl.

According to a further preferred embodiment R³ of formula I is selectedfrom the group comprising or consisting of hydrogen, C₁-C₆-alkyl, ORa,ORc, C(═O)ORa, C(═O)ORc C(═O)NRaRb, C(═O)NRaRc, C(═O)Ra, C(═O)Rc,RaC(═O)NRaRc, RaC(═O)Rb, RaC(═O)Rc, Ra(SO₂Rb), Ra(SO₂Rc), or a (═O)group (oxo).

Alternatively, R³ of formula I could also be independently from eachother selected from an aryl or a 5-6-membered heterocyclic groupcontaining at least one heteroatom selected from oxygen, nitrogen andsulfur, both the aryl, heterocyclic group being optionally substitutedby at least one C₁-C₆-alkyl, C(═O)ORa, C(═O)ORc, C₁-C₆-alkylene,trifluoromethyl, trifluoromethoxy, ORa, OC(═O)Ra, OC(═O)Rc, NRaRb,CH₂—NRaRb, NO₂, cyano, halogen, SO₂NRaRb, SO₂NRaRc, NRaSO₂Ra, NRaSO₂Rc,C₁-C₆-alkyleneC(═O)ORa, OSO₂-trifluoromethyl.

In the above set out definitions Ra and Rb are the same or different andthey are independently selected from hydrogen and C₁-C₆-alkyl, beingoptionally substituted by at least one halogen, a C₁-C₆-alkoxy or anamino group.

Furthermore, Rc does represent therein an unsubstituted or substitutedphenyl, an unsubstituted or substituted benzyl or a 3-8-memberedunsubstituted or substituted saturated 3-8-membered cyclic alkyl.

The present invention also includes the geometrical isomers, theoptically active forms, enantiomers, diastereomers of compoundsaccording to formula I, as well as their racemates. It also includes thepharmaceutically acceptable salts, e.g. hydrates, acid addition saltsthere-of, as well as the pharmaceutically active derivatives of thecarbazole derivatives of formula I. Preferred pharmaceuticallyacceptable salts of the compound I, are acid addition salts formed withpharmaceutically acceptable acids like hydrochloride, hydro-bromide,sulfate or bisulfate, phosphate or hydrogen phosphate, acetate,benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate,gluconate, methanesulfonate, benzenesulfonate, trifluoroacetate andpara-toluenesulfonate salts.

For the purpose of inhibiting Bax, the piperazine derivatives ofcarbazole according to the present invention are those compounds offormula I, wherein X is not an unsubstituted alkyl group, but a moietywhich contains at least one heteroatom, preferably between 1-6, morepreferably 1-3, and most preferred 1 or 2 heteroatoms. Preferredheteroatoms are O, F, Cl, N. The preferred methylene groups include the—CF₂-group, the —C═O group or a methylene group —CHR′, where R′ isselected of halogen, ORa, NRaRb, NHSO₂Ra, NHC(═O)NHRa, NHC(═O)ORa,NHC(═O)Ra, OC(═O)Ra, OC(═O)NHRa, OC(═O)Rc, SRa, SRc; or R′ representsCF₃. Most preferred are compounds according to formula I, wherein X is—CHF or —CHOH.

According to a preferred embodiment the substituent R² of formula I isC₁-C₆-alkylaryl (e.g. benzyl), C₁-C₆-alkylheteroaryl containing at leastone heteroatom selected from oxygen, nitrogen and sulfur, said aryl orheteroaryl or heterocyclic group being optionally substituted by atleast one C₁-C₆-alkyl, C(═O)ORa, C₁-C₄-alkylene, trifluoromethyl,trifluoromethoxy, ORa, ORc, SRa, SRc, OC(═O)Ra, OC(═O)Rc, NRaRb,CH₂—NRaRb, NO₂, cyano, halogen, SO₂NRaRb, SO₂NRaRc, NRaSO₂Ra, NRaSO₂Rc,C₁-C₄-alkylene, C(═O)ORa, OSO₂-trifluoromethyl.

According to a further particularly preferred embodiment, R² is H,benzyl or heterocyclic group.

A still further preferred embodiment consists in those piperazinederivatives of carbazole of formula I that are suitable as apharmacological tool. For such compounds of formula I, R² is afluorescent moiety. Preferred fluorescent moieties R² include(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl)acetamide,[(4,4-difluoro-5,7-dimethyl-4-bona-3a,4a-diaza-s-indacene-3-yl)methyl]acetamide,(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacen-8-yl)methyl,2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)-4-(aminoacetyl)benzoic acid,(6,7-dimethoxy-2H-chromen-2-one)-4-methyl,4,4-difluoro-1,3,5,7-tetra-methyl-4-bora-3a,4a-diaza-s-indacene-8-propionyl,4-nitro-(2,1,3-benzoxadiazol)-7-yl.

The fluorescent labelled compounds may be used to study interactionswith target proteins, cell permeability and the intracelleularlocalisation. This is useful for understanding the mode of action of theBax inhibiting compounds.

For the purpose of inhibiting Bax, particularly potent carbazolederivatives according to formula I are those wherein R⁰ and R¹ representhydrogen or lipophilic substituents, including notably, bromine,chlorine, aryl, or C₁-C₆-alkyl, preferably methyl.

The piperazine moiety within formula I could basically be substitutedwith up to 8 residues (up to 8 substituents R³), but according to apreferred embodiment R³ is hydrogen or a C₁-C₆-alkyl and most preferredis R³=hydrogen so that said piperazine moiety is attached to the centralalkyl group by its position 1 nitrogen whereas the para nitrogen(position 4) is optionally substituted by the further group R².

The most preferred Bax inhibitors according to the present invention arethose carbazole derivatives of formula I, wherein X is a —CF₂-group, a—C═O group, CH—OH, CH—F, or a methylene group —CHR′ where R′ is selectedof, alkoxy, amino or amide, R⁰ and R¹ represent hydrogen or lipophilicsubstituents, including notably, bromine, chlorine, aryl, orC₁-C₆-alkyl, preferably methyl, n and m are both 1, while o is 0 and R²is H or benzyl.

Specific examples of compounds of formula I include the following:

-   (±)-1-(4-Benzyl-piperazin-1-yl)-3-(2-methyl-carbazol-9-yl)-propan-2-ol-   (±)-4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazine-1-carboxylic    tert-butyl ester-   (±)-1-Carbazol-9-yl-3-piperazin-1-yl-propan-2-ol-   (±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylic    tert-butyl ester-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-1-(4-Benzyl-piperazin-1-yl)-3-(3,6-dibromo-carbazol-9-yl)-propan-2-ol-   (±)-1-(4-Benzyl-piperazin-1-yl)-3-carbazol-9-yl-propan-2-ol-   (±)-4-[3-(3-Bromo-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylic    tert-butyl ester-   (±)-1-(3-Bromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-1-(4-Benzyl-piperazin-1-yl)-3-(2-hydroxy-carbazol-9-yl)-propan-2-ol-   (S)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylic    tert-butyl ester-   (R)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylic    tert-butyl ester-   (S)-4-[3-(3-Bromo-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylic    tert-butyl ester-   (R)-4-[3-(3-Bromo-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylic    tert-butyl ester-   (S)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (R)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (S)-1-(3-Bromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (R)-1-(3-Bromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-1-[(3,4-Dichloro-phenyl)-piperazin-1-yl]-3-(2-methyl-carbazol-9-yl)-propan-2-ol-   (±)-1-[(3,4-Dichloro-phenyl)-piperazin-1-yl]-3-(carbazol-9-yl)-propan-2-ol-   (±)-1-[(4-Benzo[1,3]dioxol-5-yl-methyl)-piperazin-1-yl]-4-carbazol-9-yl)-propan-2-ol-   (±)-1-Carbazol-9-yl-3-[4-(4-fluoro-benzyl)-piperazin-1-yl]-propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(4-fluoro-benzyl)-piperazin-1-yl]-propan-2-ol-   (±)-1-[4-(4-Fluoro-benzyl)-piperazin-1-yl]-3-(3-phenyl-carbazol-9-yl)-propan-2-ol-   (±)-9-(2-Hydroxy-3-piperazin-1-yl-propyl)-carbazole-3,6-dicarbonitrile-   (±)-1-(3-Nitrocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-1-(3-Phenylcarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-1-(2-Hydroxycarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-1-(3,6-Diphenylcarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(3-phenyl-propyl)-piperazin-1-yl]-propan-2-ol-   (±)-1-Carbazol-9-yl-3-[4-(3-phenyl-propyl)-piperazin-1-yl]-propan-2-ol-   (±)-3,6-Dibromo-9-(2-fluoro-3-piperazin-1-yl-propyl)-carbazole-   (±)-1-(3-Amino-carbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-N-[9-(2-Hydroxy-3-piperazin-1-yl-propyl)-carbazol-3-yl]-acetamide-   (±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-2-phenoxy-ethanone-   (±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-1-phenyl-methanone-   (±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-2-(4-hydroxy-phenoxy)-ethanone-   (±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-1-(4-hydroxy-phenyl)-methanone-   (±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-1-(4-fluoro-phenyl)-methanone-   (±)-1-(4-Benzenesulfonyl-piperazin-1-yl]-3-carbazol-9-yl-propan-2-ol-   (±)-9-[3-(4-Benzyl-piperazin-1-yl)-2-methoxy-propyl]-carbazole-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-piperazin-1-yl-propan-2-one-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(2-hydroxy-3-methylamino-propyl)-piperazin-1-yl]-propan-2-ol-   1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(3-phenyl-1,2,4-thiadiazol-5-yl)-piperazin-1-yl]-propan-2-ol-   (±)-1-(4-Cyclohexylmethylpiperazin-1-yl)-3-(3,6-dibromo-carbazol-9-yl)-propan-2-ol-   (±)-1-[(4-Fluorophenyl)-piperazin-1-yl]-3-(3,6-dibromocarbazol-9-yl)-propan-2-ol-   (±)-1-(Carbazol-9-yl)-3-[4-(4-nitrobenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(Carbazol-9-yl)-3-[4-(4-methoxybenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-[4-(4-Fluorobenzyl)piperazin-1-yl]-3-{3-[4-(trifluoromethyl)phenyl]-carbazol-9-yl}propan-2-ol-   (±)-1-[4-(4-Fluorobenzyl)piperazin-1-yl]-3-{3-[4-(trifluoromethoxy)phenyl]-carbazol-9-yl}propan-2-ol-   (±)-1-(Carbazol-9-yl)-3-[4-(3-fluorobenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(Carbazol-9-yl)-3-[4-(thien-2-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(4-Butylpiperazin-1-yl)-(3-carbazol-9-yl)propan-2-ol-   (±)-4-({4-[3-carbazol-9-yl)-2-hydroxypropyl]piperazin-1-yl}methyl)phenol-   (±)-1-[4-(4-tert-Butylbenzyl)piperazin-1-yl]-3-(carbazol-9-yl)propan-2-ol-   (±)-1-(Carbazol-9-yl)-3-[4-(3,4-dichlorobenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(Carbazol-9-yl)-3-{4-[4-(methylsulfonyl)benzyl])piperazin-1-yl}propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(thien-2-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(thien-3-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(pyridin-3-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-methoxybenzl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-tert-butylbenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-trifluoromethylbenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-[4-(1,3-Benzodioxol-5-ylmethyl)piperazin-1-yl]-3-(3,6-dibromo-carbazol-9-yl)propan-2-ol-   (±)-1-(4-Cyclohexylmethylpiperazin-1-yl)-3-(3-phenylcarbazol-9-yl)-propan-2-ol-   (±)-3,6-Dibromo-9-{3-[4-(cyclohexylmethyl)piperazin-1-yl]-2-fluoropropyl}-carbazole-   (±)-9-{3-[4-(Cyclohexylmethyl)piperazin-1-yl]-2-fluoropropyl}-3-phenyl-carbazole-   (±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxypropyl]-N-(4-fluorophenyl)piperazine-1-carboxamide-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(thien-2-ylsulfonyl)piperazin-1-yl]propan-2-ol-   (±)-1-[4-(Benzylsulfonyl)piperazin-1-yl]-3-(3,6-dibromo-carbazol-9-yl)propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(3,4-dichlorobenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (±)-1-[4-(4-Fluorobenzyl)piperazin-1-yl]-3-(3-thien-2-yl-carbazol-9-yl)-propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-{4-[3,4-diethoxyphenyl)sulfonyl]piperazin-1-yl}propan-2-ol-   (±)-1-[4-(Cyclohexylmethyl)piperazin-1-yl]-3-(3,6-dichloro-carbazol-9-yl)propan-2-ol-   (±)-4-[3-(3,6-Dichlorocarbazol-9-yl)-2-hydroxypropyl]-piperazine-1-carboxylic    tert-butyl ester-   (±)-1-(3,6-Dichlorocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-oxopropyl]-piperazine-1-carboxylic    tert-butyl ester-   (±)-3,6-Dibromo-9-(2,2-difluoro-3-piperazin-1-ylpropyl)-carbazole-   (±)-4-[3-(3,6-dibromocarbazol-9-yl)-2,2-difluoropropyl]-piperazine-1-carboxylic    tert-butyl ester-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-phenylethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2,3-dihydro-1,4-benzodioxin-6-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(4-fluorobenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(3,4-dichlorobenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-{4-[4-(difluoromethoxy)benzyl]piperazin-1-yl}propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(cyclohexylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-[4-(1,3-Benzodioxol-5-ylmethyl)piperazin-1-yl]-3-(3-bromo-carbazol-9-yl)propan-2-ol-   (±)-1-(3-Bromocarbazol-9-yl)-3-[4-(4-methoxybenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromocarbazol-9-yl)-3-[4-(4-trifluoromethylbenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(3,5-dichlorobenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromocarbazol-9-yl)-3-[4-(4-tert-butylbenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-furylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-furylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-pyridin-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-Carbazol-9-yl)-3-[4-(3-pyridin-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(4-pyridin-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(quinolin-2-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-furyl-4-bromomethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(1-naphtylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-{4-[(6-Bromo-1,3-benzodioxol-5-yl)methyl]piperazin-1-yl}-3-(3-bromo-carbazol-9-yl)propan-2-ol-   (±)-1-{4-[(6-Chloro-1,3-benzodioxol-5-yl)methyl]piperazin-1-yl}-3-(3-bromo-carbazol-9-yl)propan-2-ol-   (±)-1-(3-Chlorocarbazol-9-yl)-3-[4-(4-fluorobenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Chlorocarbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(3-piperidin-1-yl-propyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Chlorocarbazol-9-yl)-3-(4-(cyclohexylpiperazin-1-yl)propan-2-ol-   (±)-1-(3-Bromocarbazol-9-yl)-3-[4-(quinolin-4-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-4-[3-(3-Chlorocarbazol-9-yl)-2-hydroxypropyl]-3,5-dimethylpiperazine-1-carboxylic    tert-butyl ester-   (±)-1-[4-(Cyclohexylmethyl)piperazin-1-yl]-3-(3,6-dichlorocarbazol-9-yl)acetone-   (±)-1-[4-(Cyclohexylmethyl)piperazin-1-yl]-3-(3,6-dibromocarbazol-9-yl)acetone-   (±)-1-(3,6-Dichlorocarbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (±)-1-[4-(Cyclohexylmethyl)piperazin-1-yl]-3-(3-phenyl    carbazol-9-yl)acetone-   (±)-1-(3-Bromocarbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Chlorocarbazol-9-yl-)-3-(3,5-dimethylpiperazine-1-yl)propan-2-ol-   (±)-1-(3-Chlorocarbazol-9-yl-)-3-(2,6-dimethylpiperazin-1-yl)propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl-)-3-piperazin-1-ylpropan-2-amine-   (±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-aminopropyl]-piperazine-1-carboxylic    tert-butyl ester-   (±)-N-Benzyl-N-[2-(3,6-dibromocarbazol-9-yl)-1-(piperazin-1-ylmethyl)ethyl]amine-   (±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-benzylaminopropyl]-piperazine-1-carboxylic    tert-butyl ester-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(2-morpholin-4-yl-2-oxoethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-{[4-(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-yl)methyl]piperazin-1-yl}propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methylacetamide)piperazin-1-yl]propan-2-ol-   (±)-1,6-Dibromocarbazol-9-yl)-3-[4-(N-(4,4-difluoro-1,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl)acetamide)piperazin-1-yl]propan-2-ol-   (±)-4-[({-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxypropyl]piperazin-1-yl}acetyl)amino]-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic    acid-   (±)-4-({-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxypropyl]piperazin-1-yl}methyl)-6,7-dimethoxy-2H-chromen-2-one-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-{4-[3-(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-yl)-propionyl]piperazin-1-yl}propan-2-ol-   (±)-1-[4-(4-Nitro-2,1,3-benzoxadiazol-7-yl)-piperazin-1-yl]-3-(3,6-dibromocarbazol-9-yl)-propan-2-ol

Thereby, the most preferred compounds are those which are selected fromthe group consisting of:

-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(4-fluoro-benzyl)-piperazin-1-yl]-propan-2-ol-   (±)-1-(3-Phenyl carbazol-9-yl)-3-piperazin-1-yl-propan-2-ol;-   (±)-3,6-Dibromo-9-(2-fluoro-3-piperazin-1-yl-propyl)-carbazole-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-piperazin-1-yl-propan-2-one-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(thien-3-ylmethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-methoxybenzyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-{4-[4-(difluoromethoxybenzyl]piperazin-1-yl}propan-2-ol-   (±)-1-(3,6-Dichlorocarbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl-)-3-piperazin-1-ylpropan-2-amine-   (R)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(4-fluoro-benzyl)-piperazin-1-yl]-propan-2-ol;-   (R)-1-(3-Phenylcarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (R)-3,6-Dibromo-9-(2-fluoro-3-piperazin-1-yl-propyl)-carbazole-   (R)-1-(3,6-Dibromo-carbazol-9-yl)-3-piperazin-1-yl-propan-2-one-   (R)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(thien-3-ylmethyl)piperazin-1-yl]propan-2-ol-   (R)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-methoxybenzyl)piperazin-1-yl]propan-2-ol-   (R)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (R)-1-(3-Bromo-carbazol-9-yl)-3-{4-[4-(difluoromethoxybenzyl]piperazin-1-yl}propan-2-ol-   (R)-1-(3,6-Dichlorocarbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (R)-1-(3,6-Dibromo-carbazol-9-yl-)-3-piperazin-1-ylpropan-2-amine-   (S)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(4-fluoro-benzyl)-piperazin-1-yl]-propan-2-ol-   (S)-1-(3-Phenylcarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (S)-3,6-Dibromo-9-(2-fluoro-3-piperazin-1-yl-propyl)-carbazole-   (S)-1-(3,6-Dibromo-carbazol-9-yl)-3-piperazin-1-yl-propan-2-one-   (S)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(thien-3-ylmethyl)piperazin-1-yl]propan-2-ol-   (S)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-methoxybenzyl)piperazin-1-yl]propan-2-ol-   (S)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (S)-1-(3-Bromo-carbazol-9-yl)-3-{4-[4-(difluoromethoxybenzyl]piperazin-1-yl}propan-2-ol-   (S)-1-(3,6-Dichlorocarbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol-   (S)-1-(3,6-Dibromo-carbazol-9-yl-)-3-piperazin-1-ylpropan-2-amine

A further aspect of the present invention is related to the use of thepiperazine derivatives of carbazole according to formula I for thepreparation of pharmaceutical compositions and their use for treatingdiseases including Alzheimer's disease, Parkinson's disease, diseasesassociated with polyglutamine tracts including Huntington's disease,spinocerebellar ataxias and dentatorubral-pallidoluysian atrophy;amyotrophic lateral, sclerosis, Crohn's disease, retinitis pigmentosaand multiple sclerosis, epilepsy), ischemia (stroke, myocardialinfarction and reperfusion injury), infertility (like prematuremenopause, ovarian failure or follicular atresia), cardiovasculardisorders (arteriosclerosis, heart failure and heart transplantation),renal hypoxia, hepatitis and AIDS.

According to a preferred embodiment, the above cited diseases or diseasestates are treated by the modulation of the Bax function, or themodulation (e.g. the inhibition) of the activation or expression of Bax,respectively, via the use of compounds of formula I, whereby the termBax function notably comprises the actually active form of Bax as anoligomer (see B. Antonsson et al. in 2000 Biochem. J., Vol. 345,271-278). Through the modulation of the Bax function, a convenientmethod of treatment of disorders mediated by Bax is expected, includingin particular neuronal disorders and/or disorders of the immune system.Said modulation could notably involve the inhibition of the activity(activation) and/or of the expression of Bax. Also, said modulation ofthe Bax function or activity could actually comprise the inhibition ordisruption for instance of the Bid interaction with Bax, which has beenshown to play a role within the context of the Bax activation leading tocytochrome c release (see J. C. Martinou et al. in 1999 The Journal ofCell Biology, 144(5), 891-901). As a result of the inhibition of the Baxactivation by Bid upon using the compounds according to formula I, thecytochrome c release could be inhibited or essentially blocked, thusproviding a convenient means to modulate the above described apoptosispathways. As a result, by said modulation of the apoptosis pathways awhole variety of disorders associated with abnormal apoptosis isexpected to be treated.

It is reported herein that the compounds of formula I are suitable to beused as a medicament, i.e. they are suitable for use in treatingdisorders of the autoimmune system and neuronal system of mammals,notably of human beings. More specifically, the compounds according toformula I, alone or in the form of a pharmaceutical composition, areuseful for the modulation, in particular for the inhibition, of the Baxfunction and/or the Bax activation. More specifically, the compoundsaccording to formula I, alone or in the form of a pharmaceuticalcomposition, are useful for the treatment or prevention of disordersassociated with abnormal expression or activation of Bax. The compoundsaccording to formula I could be employed alone or in combination withfurther pharmaceutical agents. The compounds of formula I are suitableto be used as a medicament alone or in the form of a pharmaceuticalcomposition together with suitable carriers, diluents or excipients. Thecompounds of formula I are suitable to be used for the preparation oforally administrated pharmaceutical compositions.

Thus, according to the present invention, compounds pursuant to formulaI are particularly useful for the treatment or prevention of immuno-and/or neuronal-related diseases or pathological states in whichpreferably the modulation, in particular the inhibition, of the Baxfunction and/or the Bax activation plays a crucial role, such asneurodegenerative diseases (e.g. Alzheimer's disease, Parkinson'sdisease, diseases associated with polyglutamine tracts includingHuntington's disease, spinocerebellar ataxias anddentatorubral-pallidoluysian atrophy; amyotrophic lateral sclerosis,Crohn's disease, retinitis pigmentosa and multiple sclerosis, epilepsy),ischemia (stroke, myocardial infarction and reperfusion injury),infertility (like premature menopause, ovarian failure or follicularatresia), cardiovascular disorders (arteriosclerosis, heart failure andheart transplantation), renal hypoxia, hepatitis and AIDS.

As a matter of fact, prior to the herein reported surprisingly foundpharmaceutically active pyrrolidine derivatives according to formula I,nothing was known in respect of the use of small molecule chemicalcompounds as active inhibitors of the pro-apoptosis agent Bax. Nothingwas known in respect of the possibility to disrupt or to substantiallyblock—with small molecules—the activation of Bax, for instance via Bid(being another Bcl-2 family member which is involved in the pathwaysleading to the release of cytochrome c).

A further aspect of the present invention consists in the use ofpiperazine derivatives of carbazole for the preparation of apharmaceutical composition for the treatment or prevention of disordersassociated with an abnormal Bax function or abnormal (e.g. elevated) Baxactivation, an abnormal expression or activity of Bax as well as saidpharmaceutical compositions themselves. Hence, such piperazinederivatives of carbazole useful for the preparation of a pharmaceuticalcomposition for the treatment or prevention of disorders associated withthe modulation of the Bax function or activation, in particular with theabnormal expression or activity of Bax have the above set out generalformula I. Also, the piperazine derivatives of carbazole of the presentinvention are useful for the treatment of neurodegenerative diseases(e.g. Alzheimer's disease, Parkinson's disease, diseases associated withpolyglutamine tracts including Huntington's disease, spinocerebellarataxias and dentatorubral-pallidoluysian atrophy; amyotrophic lateralsclerosis, Crohn's disease, retinitis pigmentosa and multiple sclerosis,epilepsy), ischemia (stroke, myocardial infarction and reperfusioninjury), infertility (like premature menopause, ovarian failure orfollicular atresia), cardiovascular dis-orders (arteriosclerosis, heartfailure and heart transplantation), renal hypoxia, hepatitis and AIDS.

Still a further aspect of the present invention consists in the actuallynovel carbazole derivatives of formula I, i.e. those carbazolederivatives according to formula I that have not been disclosed by theprior art. Thereby, a few compounds have been disclosed by 3 differentcompanies, i.e. by the AsInEx Company, by the Bioscreen Company and bythe Chembridge Company in as far as they have been mentioned in theircompany catalogue, without any application and most notably without anyindication concerning a potential medical use, though.

Said compounds are the following:

-   (±)-1-(4-Benzyl-piperazin-1-yl)-3-carbazol-9-yl-propan-2-ol-   (±)-1-Carbazol-9-yl-3-[4-(9H-fluoren-9-yl)-piperazin-1-yl]-propan-2-ol-   (±)-1-(4-Benzyl-piperazin-1-yl)-3-(3-chloro-carbazol-9-yl)-propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-((E)-3-phenyl-allyl)-piperazin-1-yl]-propan-2-ol-   (±)-1-(3,6-Dichloro-carbazol-9-yl)-3-[4-((E)-3-phenyl-allyl)-piperazin-1-yl]-propan-2-ol-   (±)-1-(4-Benzyl-piperazin-1-yl)-3-(3,6-dichloro-carbazol-9-yl)-propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-piperazin-1-yl-propan-2-ol-   (±)-1-Carbazol-9-yl-3-(4-pyridin-2-yl-piperazin-1-yl)-propan-2-ol-   (±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-1-phenyl-methanone-   (±)-2-{4-[3-(3,6-Dichloro-carbazol-9-yl)-2-hydroxy-propyl]-piperazin-1-yl}-ethanol-   (±)-2-[4-(3-Carbazol-9-yl)-2-hydroxy-propyl)-piperazin-1-yl]-ethanesulfonic    acid-   (±)-1-Carbazol-9-yl-3-piperazin-1-yl-propan-2-ol-   (±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(9H-fluoren-9-yl)-piperazin-1-yl]-propan-2-ol-   (±)-1-(3,6-Dichloro-carbazol-9-yl)-3-(4-pyridin-2-yl-piperazin-1-yl)-propan-2-ol-   (±)-1-Carbazol-9-yl-3-[4-(5,5-dimethyl-4,5-dihydro-thiazol-2-yl)-piperazin-1-yl]-propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(5,5-dimethyl-4,5-dihydro-thiazol-2-yl)-piperazin-1-yl]-propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-(4-thiazol-2-yl-piperazin-1-yl)-propan-2-ol-   (±)-1-Carbazol-9-yl-3-(4-thiazol-2-yl-piperazin-1-yl)-propan-2-ol-   (±)-1-(4-1,3-Benzodioxol-5-ylmethyl-piperazin-1-yl)-3-carbazol-9-yl-propan-2-ol-   (±)-1-(4-1,3-Benzodioxol-5-ylmethyl-piperazin-1-yl)-3-(3-bromo-carbazol-9-yl)-propan-2-ol-   (±)-1-(4-1,3-Benzodioxol-5-ylmethyl-piperazin-1-yl)-3-(3-chloro-carbazol-9-yl)-propan-2-ol-   (±)-1-(3-Bromo-carbazol-9-yl)-3-(4-methyl-piperazin-1-yl)-propan-2-ol-   (±)-1-(3-Chloro-carbazol-9-yl)-3-(4-methyl-piperazin-1-yl)-propan-2-ol-   (±)-1-(4-Benzyl-piperazin-1-yl)-3-(3,6-dibromo-carbazol-9-yl)-propan-2-ol

Furthermore, the following 2 compounds have been disclosed in SU-1584341of the Physico-Organic Institute in the former Soviet Union in Donetsk,

(±)-1-Carbazol-9-yl-3-piperazin-1-yl-propan-2-ol which has already beencited above as a compound disclosed in a catalogue of the AsInExcompany, and(±)-1-(4-Methyl-piperazin-1-yl)-3-carbazol-9-yl-propan-2-ol.

Said 2 compounds of SU-1584341 were described to have an anxiolyticactivity.

Hence, the entirely novel carbazole derivatives of the present inventionare those of the above set out general formula I, whereby the aboveidentified compounds are excluded. Furthermore, those compounds beingnovel in the sense of having no known medical use are those of the aboveset out general formula I, whereby the above identified 2 compounds ofSU-1584341 are excluded.

A further document disclosing piperazine derivatives of carbazole isFR-1,167,510 of July 1954. In said document, the piperazine derivativesof carbazole have the general formula V

wherein A represents a divalent, saturated aliphatic hydrocarbon radicalwith a straight or branched chain containing 2 to 6 carbon atoms.

Since within the compounds according to formula I of the presentinvention, A corresponds to a residue —(CH₂)_(n)—X—(CH₂)_(m)— wherein Xis a substituted, methylene group, more specifically a group of theformula —(CR′R″)—, whereby at least one of R′, R″ is not hydrogen, but amoiety containing at least one heteroatom, i.e. a group comprising orconsisting of a substituted C₁-C₆-alkyl, aryl or heteroaryl, aC₁-C₆-alkoxy, C₂-C₆-alkenyl, C₂-C₆-alkynyl, a primary, secondary ortertiary amino, an acylamino, aminocarbonyl, C₁-C₆-alkoxycarbonyl, acarboxylic ester acid or amide, halogen, hydroxy, sulfonyl, sulfonamide,C₁-C₆-thioalkyl, C₁-C₆-thioalkoxy or where X could be a C═O or a C═Sgroup, the above identified compounds of formula V (FR-1,167,510) aretherefore not included by the generic formula I of the presentinvention.

Also, in the South African patent No. 78/7352, a carbazole compoundaccording to the above formula V of the trade designation “Rimcazole”(an anxiolytic agent), where A is —(CH₂)₃— and whereby the piperazinemoiety is di-substituted with a methyl group is disclosed. Furthercompounds according to formula V that do have a non-substituted alkylbridge between the piperazine and the carbazole moiety, wherein R is adiphenylmethyl or an ethylester radical or hydrogen are described byKomissarenko et al. in Khim.-Farm. Zh, 24(10), 54-56; 1990 as well asHarfenist et al. in J. Org. Chem., 50(9), 1356; 1985. Such compounds areequally not at all included by the formula I.

The compounds of formula I may contain one or more asymmetric centersand may therefore exist as enantiomers or diasteroisomers. It is to beunderstood that the invention includes both mixtures and separateindividual isomers or enantiomers of the compounds of formula I. In aparticularly preferred embodiment the carbazole derivatives according toformula I are obtained in an enantiomeric excess of at least 52% ee,preferably of at least 92-98% ee.

A further aspect of the present invention consists in the use ofcarbazole derivatives for the preparation of a pharmaceuticalcomposition for the treatment or prevention of disorders associated withabnormal Bax function or Bax activation, an abnormal expression oractivity of Bax as well as said pharmaceutical compositions themselves.Such a composition could be prepared by using both the novel and knowncompounds according to formula I including those of FR-1,167,510according to formula V as well as those of SU-1584341 and South Africanpatent No. 78/7352. Hence, such piperazine derivatives of carbazoleuseful for the preparation of a pharmaceutical composition for thetreatment or prevention of disorders associated with the modulation ofthe Bax function or activation, in particular with the abnormalexpression or activity of Bax have the general formula

as well as the pharmaceutically acceptable salts and thereof, in racemicor essentially enantiomeric pure form, wherein R⁰, R¹, R², R³, k, l, m,n and o as well as X are as above defined, i.e. according to the generaldefinition of formula I.

Still a further object of the present invention is the use of carbazolecompounds of formula I, wherein R² is a fluorescent moiety, as apharmacological tool. Thereby, the preferred fluorescent moieties R²include(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl)acetamide,[(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl]acetamide,(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacen-8-yl)methyl,2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)-4-(aminoacetyl)benzoic acid,(6,7-dimethoxy-2H-chromen-2-one)-4-methyl,4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-propionyl,4-nitro-(2,1,3-benzoxadiazol)-7-yl. The fluorescent labelled compoundscan be used to study interactions with target proteins, cellpermeability and the intracelleular localisation. This is helpful forunderstanding the mode of action of the Bax inhibiting compounds.

Still a further object of the present invention is a process forpreparing the novel piperazine derivatives of carbazole according toformula I which have been set out above.

The piperazine derivatives of carbazole of this invention can beprepared from readily available starting materials using the followinggeneral methods and procedures.

It will be appreciated that where typical or preferred experimentalconditions (i.e. reaction temperatures, time, moles of reagents,solvents, etc.) are given, other experimental conditions can also beused unless otherwise stated. Optimum reaction conditions may vary withthe particular reactants or solvent used, but such conditions can bedetermined by a person skilled in the art by routine optimisationprocedures.

Generally, the piperazine derivatives of the carbazole compoundaccording to the general formula I could be obtained following toProtocol A, i.e. by reacting a carbazole derivative of formula II

whereby the substituents R⁰, R¹, k and l are as above defined, with apiperazine derivative according to the general formula

whereby R² and R³, m, n and o are as above defined, while Y could be anyappropriate leaving group. Particularly preferred leaving groups arethose selected from the group comprising or consisting of halogen, analiphatic or aromatic sulfonyloxy group such as methanesulfonyloxy or4-toluenesulfonyloxy or 3-nitro-benzenesulfonyloxy group.

A particularly preferred method to prepare compounds of formula I inwhich m=1 and X=CH(OH), is the condensation of compound of formula IIand the compound of formula IV.

The reaction is carried out between compound III or IV and an anion ofcompound H which can be formed by reaction of compound II and a basesuch as sodium hydride, n-butyl lithium in a suitable organic solvent,such as dimethylformamide (DMF) or tetrahydrofuran (THF) for severalhours, e.g. 2 hours to 16 hours at room temperature or e.g. at 60° C.

Compounds of formula I can be prepared as individual enantiomers or inan enantiomeric enriched form from the appropriate enantiomer of formulaIII or as a racemic mixture from the appropriate racemic compound offormula III. Individual enantiomers of the invention can be preparedfrom racemates by resolution using methods known in the art for theseparation of racemic mixtures into their constituent enantiomers, forexample using HPLC on a chiral column such as Chiralpak AD, ChiralcelOJ, or using separation of salts of diastereomers.

Compounds of formula II are commercially available compounds or preparedby standard synthetic techniques as hereinafter described in theExamples.

Compounds of formula III can be prepared from the exposure of thecorresponding piperazine compounds of formula VI and compounds offormula VIII:

In which LG is any good leaving group suitably e.g. halogen such aschlorine, bromine or iodine, or an aliphatic or aromatic sulfonyloxygroup such as methanesulfonyloxy or 4-toluenesulfonyloxy or3-nitro-benzenesulfonyloxy or 4-nitro-benzenesulfonyloxy group. X is aspreviously defined or can be protected by any suitable protecting group.Y′ is a precursor of Y, e.g. a protected form of an alcohol, which,after the condensation with the piperazine compound of formula VI, willbe transformed into Y, according to procedures known by a person skilledin the art. For all the protection, de-protection methods, see Philip J.Kocienski, in “Protecting Groups”, Georg Thieme Verlag Stuttgart, NewYork, 1994 and, Theodora W. Greene and Peter G. M. Wuts in “ProtectiveGroups in Organic Synthesis”, Wiley-Interscience, 1991.

The reaction provides racemic compounds of formula III. (R) as well as(S) enantiomers can be obtained depending upon whether (R) or (S)compound of formula VIII was used as the starting material.

Compounds of formula IV can be prepared from the exposure of thecorresponding piperazine compound VI to the compound of formula VIIcontaining any good leaving group (LG) suitably e.g. halogen such asbromine or iodine, or an aliphatic or aromatic sulfonyl-oxy group suchas methanesulfonyloxy or 4-toluenesulfonyloxy or3-nitrobenzenesulfonyloxy or 4-nitro-benzenesulfonyloxy group in thepresence of a base such as potassium carbonate in a suitable solvente.g. acetonitrile.

The reaction provides racemic compounds of formula IV. (R) as well as(S) enantiomers can be obtained depending upon whether (R) or (S)compound of formula VII was used as the starting material.

Compounds of formula VI, VII and VIII are commercially available orprepared by standard synthetic techniques as hereinafter described inthe examples.

An alternative method of preparation of the carbazole compounds offormula I consists in the following Protocol B. Thereby, the piperazinederivatives of the carbazole compound according to the general formula Icould be obtained by reacting a carbazole derivative of formula IX

whereby the substituents R⁰, R¹, k, l, m and n are as above defined,while Y could be any appropriate leaving group with a piperazinederivative according to the general formula VI. Particularly preferredleaving groups are those selected from the group comprising orconsisting of halogen, an aliphatic or aromatic sulfonyloxy group suchas methanesulfonyloxy or 4-toluenesulfonyloxy or3-nitro-benzenesulfonyloxy group.

The preferred method to prepare compounds of formula I in which m=1 andX=CH(OH), is the condensation of compound of formula VI and the compoundof formula X.

The reaction is carried out between compound I× or XIII and an compoundVI in a suitable organic solvent, such as ethanol (EtOH), acetonitrile(ACN) or a mixture of EtOH/tetrahydrofuran (THF) (1/1) for severalhours, e.g. 2 hours to 16 hours at room temperature or at 60° C. or at80° C.

Compounds of formula I can be prepared as individual enantiomers or inan enantiomeric enriched form from the appropriate enantiomer of formulaXIII or as a racemic mixture from the appropriate racemic compound offormula X. Individual enantiomers of the invention can be prepared fromracemates by resolution using methods known in the art for thesepa-ration of racemic mixtures into their constituent enantiomers, forexample using HPLC on a chiral column such as Chiralpak AD, ChiralcelOJ, or using separation of salts of diastereo-mers.

Compounds of formula IX can be prepared from the exposure of thecorresponding carbazole compounds of formula II and compounds of formulaXI:

In which LG is any suitable leaving group including halogen such aschlorine, bromine or iodine, or an aliphatic or aromatic sulfonyloxygroup such as methanesulfonyloxy or 4-toluenesulfonyloxy or3-nitro-benzenesulfonyloxy or 4-nitro-benzenesulfonyloxy group. X is aspreviously defined or can be protected by any suitable protecting group.Y′ is a precursor of Y, e.g. a protected form of an alcohol, which,after the condensation with the carbazole compound of formula II, willbe transformed into Y, according to procedures known by a person skilledin the art. For all the protection, de-protection methods, see Philip J.Kocienski, in “Protecting Groups”, Georg Thieme Verlag Stuttgart, NewYork, 1994 and, Theodora W. Greene and Peter G. M. Wuts in “ProtectiveGroups in Organic Synthesis”, Wiley-Interscience, 1991.

The reaction provides racemic compounds of formula IX. (R) as well as(S) enantiomers can be obtained depending upon whether (R) or (S)compound of formula XII was used as the starting material.

Compounds of formula X may be prepared by contacting the correspondingcarbazole compound H with the compound of formula XII containing asuitable leaving group (LG) including for instance halogen such asbromine or iodine, or an aliphatic or aromatic sulfonyl-oxy group suchas methanesulfonyloxy or 4-toluenesulfonyloxy or3-nitro-benzenesulfonyl-oxy or 4-nitro-benzenesulfonyloxy group in thepresence of a base such as sodium hydride in a suitable solvent e.g. THFor acetonitrile.

The reaction provides racemic compounds of formula XIII (R) as well as(S) enantiomers can be obtained depending upon whether (R) or (S)compound of formula XII was used as the starting material.

Compound of formula XI and XII are commercially available compounds ormay be prepared upon using standard synthetic techniques as hereinafterdescribed in the examples.

A more preferred preparation of compound XIII is the exposure of thecorresponding carbazole compound II to the compound of formula XII whenLG is an hydroxy group. The reaction is carried out by standardMitsunobu reaction conditions i.e. diethyl (E)-1,2-diazenedicarboxylate(DEAD), PPh₃ in anhydrous THF or using(E)-N¹,N¹,N²,N²-tetramethyl-1,2-diazenedicarboxamide (TMAD) and PBu₃, or(trimethyl phosphoranylidene)acetonitrile (CMMP) which is preparedaccording to the protocol of Tetsuto, Tsunoda Tett. Lett. 1996, 37(14),2459-2462, or (tributylphosphoranylidene) acetonitrile (CMBP) in asuitable anhydrous solvent such as toluene or THF.

Following a third approach (Protocol C), a one-step-procedure, thepiperazine derivatives of carbazole compound according to the generalformula I could be obtained by reacting the carbazole derivative offormula II, the compound of formula VII containing any good leavinggroup suitably e.g. halogen such as bromine or iodine, or an aliphaticor aromatic sulfonyloxy group such as methanesulfonyloxy or4-toluenesulfonyloxy or 3-nitrobenzenesulfonyloxy or4-nitrobenzenesulfonyloxy group, e.g. (±)-3-nitrobenzene sulfonic acidoxiranylmethyl ester, and the piperazine derivative of formula VI. Thereaction is carried out in a basic medium, such as ethanol, THF or ACNin the presence of a suitable base e.g. NaH.

The method of preparation of the carbazole compounds of formula Iaccording to Protocole B and C has the specific advantage of being moreconvenient and economic, in the sense that it comprises less steps.

Compounds of formula XI and XII are commercially available or preparedby synthetic techniques as hereinafter described in Examples.

Particularly preferred intermediate compounds of formula II, III, IV, IXand XIII for the pre-paration of a compound according to formula I areselected from the group consisting of:

-   (t)-1-Benzyl-4-oxiranylmethyl-piperazine-   (s)-4-Oxiranylmethyl-piperazine-1-carboxylic acid tert-butyl ester-   (R)-4-Oxiranylmethyl-piperazine-1-carboxylic acid tert-butyl ester-   (±)-1-(3,4-Dichloro-phenyl)-4-oxiranylmethyl-piperazine-   (±)-1-(1,3-Benzodioxol-5-ylmethyl)-4-oxiranylmethyl-piperazine-   (±)-1-(4-Fluoro-benzyl)-4-oxiranylmethyl-piperazine-   3-Phenyl-9H-carbazole-   3,6-Diphenyl-9H-carbazole-   (±)-1-Oxiranylmethyl-4-(3-phenyl-propyl)-piperazine-   (±)-1-Oxiranylmethyl-4-(3-phenyl-1,2,4-thiadiazol-5-yl)-piperazine-   (±)-1-(Cyclohexylmethyl)-4-(oxiran-2-ylmethyl)piperazine-   (±)-1-(4-Fluorophenyl)-4-oxiran-2-ylmethyl)piperazine-   3-[4-(Trifluoromethyl)phenyl]-9H-carbazole-   3-[4-(Trifluoromethoxy)phenyl]-9H-carbazole-   (±)-3,6-Dibromo-9-(oxiran-2-ylmethyl)carbazole-   3-Thien-2-yl-9H-carbazole-   (±)-3-Chloro-9-(oxiran-2-ylmethyl)carbazole-   (±)-1-(Oxiran-2-ylmethyl)-4-(3-piperidin-1-ylpropyl)piperazine-   (±)-3-Nitro-9-(oxiran-2-ylmethyl)carbazole-   3-Nitro-9-[(1E)-3-(3-nitrocarbazol-9-yl)propen-1-enyl]carbazole.

The above mentioned novel intermediate compounds are a further aspect ofthe present invention.

According to a further general process, compounds of formula I can beconverted to alternative compounds of formula I, employing suitableinterconversion techniques such as hereinafter described in theExamples.

If the above set out general synthetic methods are not applicable forobtaining compounds according to formula I and/or necessaryintermediates for the synthesis of compounds of formula I, suitablemethods of preparation known by a person skilled on the art should beused. In general, the synthesis pathways for any individual compound offormula I will depend on the specific substitutents of each molecule andupon the ready availability of intermediates necessary; again suchfactors being appreciated by those of ordinary skill in the art.

Compounds of this invention can be isolated in association with solventmolecules by crystallization from evaporation of an appropriate solvent.The pharmaceutically acceptable acid addition salts of the compounds offormula I, which contain a basic center, may be prepared in aconventional manner. For example, a solution of the free base may betreated with a suitable acid, either neat or in a suitable solution, andthe resulting salt isolated either by filtration or by evaporation undervacuum of the reaction solvent. Pharmaceutically acceptable baseaddition salts may be obtained in an analogous manner by treating asolution of compound of formula I with a suitable base. Both types ofsalt may be formed or interconverted using ion-exchange resintechniques.

A final aspect of the present invention is related to the formulationscontaining the active compounds according to formula I. When employed aspharmaceuticals, the carbazole derivatives of the present invention aretypically administered in the form of a pharmaceutical composition.Hence, pharmaceutical compositions comprising a compound of formula Iand a pharmaceutically acceptable carrier, diluent or excipienttherefore are also within the scope of the present invention. A personskilled in the art is aware of a whole variety of such carrier, diluentor excipient compounds suitable to formulate a pharmaceuticalcomposition. Also, the present invention provides compounds for use as amedicament. In particular, the invention provides the compounds offormula I for use as Bax antagonist, for the treatment of disorders ofmammals, notably of human beings associated with inappropriate celldeath, including neurodegenerative disorders, diseases associated withpolyglutamine tracts, epilepsy, ischemia, infertility, cardiovasculardisorders, renal hypoxia, hepatitis and AIDS, either alone or incombination with other medicaments.

The compounds of the invention, together with a conventionally employedadjuvant, carrier, diluent or excipient may be placed into the form ofpharmaceutical compositions and unit dosages thereof, and in such formmay be employed as solids, such as tablets or filled capsules, orliquids such as solutions, suspensions, emulsions, elixirs, or capsulesfilled with the same, all for oral use, or in the form of sterileinjectable solutions for parenteral administration (includingsubcutaneous use). Such pharmaceutical compositions and unit dosageforms thereof may comprise ingredients in conventional proportions, withor without additional active compounds or principles, and such unitdosage forms may contain any suitable effective amount of the activeingredient commensurate with the intended daily dosage range to beemployed.

When employed as pharmaceuticals, the carbazole derivatives of thisinvention are typically administered in the form of a pharmaceuticalcomposition. Such compositions can be prepared in a manner well known inthe pharmaceutical art and comprise at least one active compound.Generally, the compounds of this invention are administered in apharmaceutically effective amount. The amount of the compound actuallyadministered will typically be determined by a physician, in the lightof the relevant circumstances, including the condition to be treated,the chosen route of administration, the actual compound administered,the age, weight, and response of the individual patient, the severity ofthe patient's symptoms, and the like.

The pharmaceutical compositions of these inventions can be administeredby a variety of routes including oral, rectal, transdermal,subcutaneous, intravenous, intramuscular, and intranasal. Depending onthe intended route of delivery, the compounds are preferably formulatedas either injectable or oral compositions. The compositions for oraladministration can take the form of bulk liquid solutions orsuspensions, or bulk powders. More commonly, however, the compositionsare presented in unit dosage forms to facilitate accurate dosing. Theterm “unit dosage forms” refers to physically discrete units suitable asunitary dosages for human subjects and other mammals, each unitcontaining a predetermined quantity of active material calculated toproduce the desired therapeutic effect, in association with a suitablepharmaceutical excipient. Typical unit dosage forms include prefilled,premeasured ampoules or syringes of the liquid compositions or pills,tablets, capsules or the like in the case of solid compositions. In suchcompositions, the carbazole compound is usually a minor component (fromabout 0.1 to about 50% by weight or preferably from about 1 to about 40%by weight) with the remainder being various vehicles or carriers andprocessing aids helpful for forming the desired dosing form.

Liquid forms suitable for oral administration may include a suitableaqueous or nonaqueous vehicle with buffers, suspending and dispensingagents, colorants, flavors and the like.

Solid forms may include, for example, any of the following ingredients,or compounds of a similar nature: a binder such as microcrystallinecellulose, gum tragacanth or gelatine; an excipient such as starch orlactose, a disintegrating agent such as alginic acid, Primogel, or cornstarch; a lubricant such as magnesium stearate; a glidant such ascolloidal silicon dioxide; a sweetening agent such as sucrose orsaccharin; or a flavoring agent such as peppermint, methyl salicylate,or orange flavoring.

Injectable compositions are typically based upon injectable sterilesaline or phosphate-buffered saline or other injectable carriers knownin the art. As above mentioned, the carbazole compound or carbazolecompounds of formula I in such compositions is/are typically a minorcomponent, frequently ranging between 0.05 to 10% by weight with theremainder being the injectable carrier and the like.

The above described components for orally administered or injectablecompositions are merely representative. Further materials as well asprocessing techniques and the like are set out in Part 8 of Remington'sPharmaceutical Sciences, 17^(th) Edition, 1985, Marck PublishingCompany, Easton, Pa., which is incorporated herein be reference.

The compounds of this invention can also be administered in sustainedrelease forms or from sustained release drug delivery systems. Adescription of representative sustained release materials can also befound in the incorporated materials in Remington's PharmaceuticalSciences.

In the following the present invention shall be illustrated by means ofsome examples which are not construed to be viewed as limiting the scopeof the invention.

EXAMPLES

The following abbreviations are hereinafter used in the accompanyingexamples: min (minute), hr (hour), g (gram), mmol m.p. (melting point),eq (equivalents), mL (milliliter), μL (microliters), mL (milliliters),DCM (dichloromethane), TFA (trifluoro-acetic acid), rt (roomtemperature), DMSO (dimethylsulfoxide), DMSO-d₆ (deuterateddimethylsulfoxide), THF (tetrahydrofuran), MgSO₄ (magnesium sulfate),CDCl₃ (deuterated chloroform), TEA (triethyl amine), NaH (sodiumhydride), nBuLi (n Butyl Lithium), EtOAc (ethyl acetate), cHex(cyclohexanes), Et₂O (diethyl ether), ACN (acetonitrile), PetEther(petroleum ether), NaHCO₃ (sodium bicarbonate), HOBt(1-hydroxybenzotriazole), EDCI(1-(3-dimethyl-amino-propyl)-3-ethylcarbodiimide), dimethylformamide(DMF), K₂CO₃ (potassium carbonate), TMOF (trimethyl ortho formiate),DAST [(diethylamino)sulfur trifluoride], DEAD (diethyl(E)-1,2-diazenedicarboxylate), CMMP (trimethylphosphoranylidene)acetonitrile.

The below described intermediates 1-19 are novel. They are particularlyadvantageous for the preparation of the compounds according to formulaI, notably for the hereinafter exemplified compounds 1-124.

Intermediate 1 (±)-1-Benzyl-4-oxiranylmethyl-piperazine

To a solution of 2-bromomethyl-oxirane (2.3 mL, 27.5 mmol) in 40 mL ofACN are added K₂CO₃ (3.8 g, 27.5 mmol) and a solution of1-benzyl-piperazine (5.81 g, 25 mmol) in 60 mL of ACN. After 24 hr at rtthe reaction is judged to be completed by tlc monitoring (SiO₂, DCM:MeOH20:1). The reaction mixture is filtered off and the filtrate isconcentrated in vacuo. Flash chromatography on a 7×15 cm² column of SiO₂using DCM:MeOH (20:1) as eluting solvent and removal of the solventgives the title compound (4.87 g, 84%) as a colorless oil.

¹H NMR (CDCl₃, 300 MHz) δ 7.34-7.24 (m, 5H), 3.55 (system AB, 2H,Aa=14.9 Hz, J=13.3 Hz), 3.11 (m, 1H), 2.76 (dd, 1H, J=9.1, 5.1 Hz), 2.70(dd, 1H, J=13.2, 6.6 Hz), 2.53-2.47 (m, 8H), 2.48 (dd, 1H, J=5.1, 2.7Hz), 2.31 (dd, 1H, J=13.2, 6.6 Hz).

Intermediate 2 (S)-4-Oxiranylmethyl-piperazine-1-carboxylic acidtert-butyl ester

The same method as employed in the preparation of Intermediate 1 butstarting from piperazine-1-carboxylic acid tert-butyl ester and(R)-(−)-3-nitro-benzenesulfonic acid oxiranylmethyl ester gives afterflash chromatography the title compound as a yellow oil in a 70% yield.

¹H NMR (CDCl₃, 300 MHz) δ 3.42 (m, 3H), 3.08 (m, 1H), 2.76 (m, 2H),2.57-2.45 (m, 6H), 2.25 (dd, 1H, J=13.3, 6.9 Hz), 1.44 (s, 9H). 96% ee(Chiralcel OJ column, rt, hexanes:iPrOH (95:5), 210 nM, 0.5 mL/min).

Intermediate 3 (R)-4-Oxiranylmethyl-piperazine-1-carboxylic acidtert-butyl ester

The same method as employed in the preparation of Intermediate 1 butstarting from piperazine-1-carboxylic acid tert-butyl ester and(S)-(+)-3-nitro-benzenesulfonic acid oxiranylmethyl ester gives afterflash chromatography the title compound as a yellow oil in a 68% yield.

¹H NMR (CDCl₃, 300 MHz) δ 3.42 (m, 3H), 3.08 (m, 1H), 2.76 (m, 2H),2.57-2.45 (m, 6H), 2.25 (dd, 1H, J=13.3, 6.9 Hz), 1.44 (s, 9H). 96% ee(Chiralcel OJ column, rt, hexanes:iPrOH (95:5), 210 nM, 0.5 mL/min).

Intermediate 4 (±)-1-(3,4-Dichloro-phenyl)-4-oxiranylmethyl-piperazine

The same method as employed in the preparation of Intermediate 1 butstarting from 1-(3,4-dichloro-phenyl)-piperazine) and3-nitro-benzenesulfonic acid oxiranylmethyl ester gives after Biotagechromatography the title compound as a yellow oil in a 97% yield.

¹H NMR (CDCl₃, 300 MHz) δ 725 (d, 1H, J=9.0 Hz), 6.93 (d, 1H, J=2.9 Hz),6.71 (dd, 1H, J=9.0, 2.9 Hz), 3.17 (m, 4H), 2.84 (m, 1H), 2.79 (dd, 1H,J=13.3, 3.2 Hz), 2.78-2.62 (m, 5H), 2.48 (dd, 1H, J=5.0, 2.7 Hz), 2.26(dd, 1H, J=13.3, 7.0 Hz).

Intermediate 5(±)-1-(1,3-Benzodioxol-5-ylmethyl)-4-oxiranylmethyl-piperazine

The same method as employed in the preparation of Intermediate 1 butstarting from 1-benzo[1,3]dioxo-5-ylmethylpiperazine gives after flashchromatography the title compound as an orange oil in a 100% yield.

¹H NMR (DMSO-d₆, 300 MHz) δ 6.82 (m, 2H), 6.72 (dd, 1H, J=7.8, 1.5 Hz),5.97 (s, 2H), 4.07 (dd, 1H, J=5.4, 5.4 Hz), 3.32 (s, 2H), 2.97 (m, 1H),2.66 (dd, 1H, J=3.6, 1.3 Hz), 2.57 (dd, 1H, J=13.2, 3.6 Hz), 2.41 (dd,1H, J=5.1, 2.7 Hz), 2.34 (br s, 8H), 2.17 (dd, 1H, J=13.2, 6.6 Hz).

Intermediate 6 (±)-1-(4-Fluoro-benzyl)-4-oxiranylmethyl-piperazine

The same method as employed in the preparation of Intermediate 1 butstarting from 1-(3-fluoro-benzyl)-piperazine gives after flashchromatography the title compound as a colorless oil in a 83% yield.

¹H NMR (CDCl₃, 300 MHz) δ 7.25 (m, 2H), 6.95 (m, 2H), 3.45 (system AB,2H, Δα=14.9, J=13.3 Hz), 3.07 (m, 1H), 2.70 (dd, 1H, J=13.2, 3.6 Hz),2.60-2.44 (m, 10H), 2.26 (dd, 1H, J=13.2, 6.7 Hz).

Intermediate 7 3-Phenyl-9H-carbazole

To a solution of 3-bromo-9H-carbazole (0.4 g, 1.6 mmol) in DMF:H₂O (15mL:2 mL) are added tetrakis(triphenylphosphine)-palladium (0.184 g, 0.1eq), potassium carbonate (0.66 g, 3 eq) and phenyl boronic acid (0.58 g,3 eq). The resulting mixture is stirred at 80° C. for 4 hr. Aftercooling to rt, DCM (10 mL) is added and the reaction mixture is quenchedwith brine. After extraction, drying over MgSO₄ and concentration invacuo, the residue is purified via flash chromatography usingPetEther:EtOAc 10:1 as eluting solvent to give the title compound (0.35g, 1.44 mmol) as a white powder in a 90% yield. M.p.: 221° C.

¹H NMR (CDCl₃, 300 MHz) δ 8.17 (d, 1H, J=1.5 Hz), 8.01 (d, 1H, J=7.8Hz), 7.97 (br s, 1H), 7.81-7.37 (m, 3H), 7.60-7.30 (m, 7H).

Intermediate 8 3,6-Diphenyl-9H-carbazole

The same method as employed in the preparation of Intermediate 7 butstarting from 3,6-dibromo-9H-carbazole gives after flash chromatographythe title compound as a white solid in a 33% yield. M.p.: 145-147° C.

¹H NMR (CDCl₃, 300 MHz) δ 8.32 (d, 2H, J=1.2 Hz), 8.09 (s, 1H),7.72-7.66 (m, 6H), 7.50-7.43 (m, 6H), 7.35-7.30 (m, 2H).

Intermediate 9 (±)-1-Oxiranylmethyl-4-(3-phenyl-propyl)-piperazine

The same method as employed in the preparation of Intermediate 1 butstarting from 1-(3-phenylpropyl)-piperazine and 3-nitro-benzenesulfonicacid oxiranylmethyl ester gives after Biotage chromatography the titlecompound as a yellow oil in a 80% yield.

¹H NMR (CDCl₃, 300 MHz) δ 7.23-7.09 (m, 5H), 3.02 (m, 1H), 2.72-2.32 (m,15H), 2.26 (dd, 1H, J=13.2, 6.8 Hz), 1.82 (m, 2H).

Intermediate 10(±)-1-Oxiranylmethyl-4-(3-phenyl-1,2,4-thiadiazol-5-yl)-piperazine

The same method as employed in the preparation of Intermediate 1 butstarting from 1-(3-phenyl-1,2,4-thiadiazol-5-yl)-piperazine and3-nitro-benzenesulfonic acid oxiranylmethyl ester gives after Biotagechromatography the title compound as a white powder in a 60% yield.

¹H NMR (CDCl₃, 300 MHz) δ 8.07 (m, 2H), 7.30 (m, 3H), 3.53 (br s, 3H),3.14-2.20 (m, 10H).

Intermediate 11 (±)-1-(Cyclohexylmethyl)-4-(oxiran-2-ylmethyl)piperazine

The same method as employed in the preparation of Intermediate 1 butstarting from 1-cyclohexylmethyl piperazine and(±)-3-nitro-benzenesulfonic acid oxiranylmethyl ester gives after flashchromatography the title compound as a yellow oil in a 92% yield.

¹H NMR (CDCl₃, 300 MHz) δ 3.02 (m, 1H), 2.72-2.32 (m, 10H), 2.26 (dd,1H, J=13.2, 6.8 Hz), 2.06 (d, 1H, J=7.2 Hz), 1.82 (m, 6H), 1.4 (m, 1H),1.4 (m, 4H), 0.8 (m, 2H).

Intermediate 12 (±)-1-(4-Fluorophenyl)-4-oxiran-2-ylmethyl)piperazine

The same method as employed in the preparation of Intermediate 1 butstarting from 4-fluorophenylpiperazine, dihydrochloride and(±)-3-nitro-benzenesulfonic acid oxiranyl-methyl ester gives after flashchromatography the title compound as a yellow oil in a 25% yield.

¹H NMR (CDCl₃, 300 MHz) δ 6.97-6.83 (2 d, J=8.3 Hz, 4H), 3.2 (m, 5H),2.80-2.60 (m, 8H), 2.50 (dd, 1H, J=5.0, 2.7 Hz), 2.3 (dd, 1H, J=13.2,6.9 Hz).

Intermediate 13 3-[4-(Trifluoromethyl)phenyl]-9H-carbazole

The same method as employed in the preparation of Intermediate 7 butstarting from 4-trifluoromethylphenyl boronic acid gives after Biotagechromatography the title compound as a white powder in a 60% yield.

FI-MS (APCI): m/z observed in a negative mode: 310.0.

Intermediate 14 3-[4-(Trifluoromethoxy)phenyl]-9H-carbazole

The same method as employed in the preparation of Intermediate 7 butstarting from 4-trifluoromethoxyphenyl boronic acid gives after Biotagechromatography the title compound as a white powder in a 97% yield.

FI-MS (APCI): m/z observed in a negative mode: 325.8.

Intermediate 15 (±)-3,6-Dibromo-9-(oxiran-2-ylmethyl)carbazole

Following Protocol A:

At rt a solution of 3,6-dibromo-9H-carbazole (5.23 g, 16.10 mmol) inanhydrous THF (200 mL) is treated with NaH (0.703 g, 16.10 mmol, 55% inmineral oil) and a solution of (±)-3-nitro-benzene sulfonic acidoxiranyl methyl ester (4.17 g, 16.10 mmol) in anhydrous THF (30 mL).After 16 hours of stirring at rt the reaction mixture is quenched with asaturated aqueous solution of sodium hydrogenocarbonate (200 mL).Extraction with Et₂O (500 mL+2×250 mL), drying over MgSO₄ andevaporation under reduced pressure gives a yellow solid. Flashchromatography on a 7×27 cm² column of SiO₂ using DCM/Petroleum Ether(1/1) gives the title compound (3.65 g, 9.58 mmol) as a white solid in a59% yield.

Following Protocol B:

At rt under an inert atmosphere, (±)-glycyclol (0.129 g, 1.74 mmol, 120uL) is added into a solution of 3,6-dibromo-9-H-carbazole (0.565 g, 1.74mmol) in anhydrous toluene (10 mL). Then CMMP (which is preparedaccording to the protocol of Tetsuto, Tsunoda Tett. Lett. 1996, 37(14),2459-2462) (0.20 g, 1.74 mmol) is added. The resulting mixture isallowed to stir at rt for 16 hours. The solvent is evaporated and thecrude compound is purified via flash chromatography using Biotage deviceand Petroleum Ether/DCM (3/1) as eluant to give the title compound in a60% yield as a white solid and 5% of(±)-3,6-dibromo-9-[(1E)-3-(3,6-dibromocarabazol-9-yl)propen-1-enyl]carbazoleas a grey powder.

-   (±)-3,6-Dibromo-9-(oxiran-2-ylmethyl)carbazole:

¹H NMR (CDCl₃, 300 MHz) δ 8.11 (d, 2H, J=1.9 Hz), 7.55 (dd, 2H, J=8.7,1.9 Hz), 7.31 (d, 2H, J=8.7 Hz), 4.62 (dd, J=16.0, 2.7 Hz, 1H), 4.25(dd, J=16.0, 5.1 Hz, 1H), 3.29 (dddd, J=5.1, 4.0, 2.7, 2.6 Hz, 1H), 2.79(dd, J=4.7, 4.0 Hz, 1H), 2.47 (dd, 1H, J=4.7, 2.6 Hz, 1H).

-   (±)-3,6-Dibromo-9-[(1E)-3-(3,6-dibromocarabazol-9-yl)propen-1-enyl]carbazole:

¹H NMR (CDCl₃, 300 MHz) δ 8.6-8.4 (4H, m), 8.07-7.41 (9H, m), 6.34-6.1(1H, m), 5.5-5.2 (2H, d, J=6.0 Hz).

FI-MS (APCI): m/z observed in a negative mode: 687.2

Intermediate 16 3-Thien-2-yl-9H-carbazole

Under an inert atmosphere a solution of 3-bromo-2-yl-9H-carbazole (0.100g, 0.408 mmol) in degassed NMP (10 mL) is treated with triphenyl arsine(0.025 g, 0.0816 mmol, 0.2 equiv.), Pd₂ dba₃ (0.0187 g, 0.0204 mmol, 5mol %) and tributyl(thien-2-yl)stannane (0.167 mg, 0.449 mmol, 1.1equiv.). The resulting mixture is allowed to stir for 20 hours at 80° C.The reaction mixture is quenched with a solution of KF (1M) for 1 hour.Extraction with DCM, drying over MgSO₄ and evaporation under reducedpressure gives a residue. HPLC purification and recrystallization fromEt₂O gives the title compound as a yellow solid in a 20% yield.

FI-MS (APCI): m/z observed in a negative mode: 248.0.

Intermediate 17 (±)-3-Chloro-9-(oxiran-2-ylmethyl)carbazole

The same method as employed in the preparation of Intermediate 15 butstarting from 3-chloro-9H-carbazole gives after Biotage chromatographythe title compound as a yellow oil in a 78% yield.

¹H NMR (CDCl₃, 300 MHz) δ 8.03 (m, 1H), 7.49-7.25 (m, 6H), 4.62 (dd, 1H,J=16.0, 2.7 Hz), 4.25 (dd, 1H, J=16.0, 5.1 Hz), 3.29 (dddd, 1H, J=5.1,4.0, 2.7, 2.6 Hz), 2.79 (dd, 1H, J=4.7, 4.0 Hz), 2.47 (d, 1H, J=4.7, 2.6Hz).

Intermediate 18(±)-1-(Oxiran-2-ylmethyl)-4-(3-piperidin-1-ylpropyl)piperazine

The same method as employed in the preparation of Intermediate 1 butstarting from 1-(3-piperidin-1-ylpropyl)piperazine and(±)-3-nitro-benzenesulfonic acid oxiranylmethyl ester gives after flashchromatography the title compound as an orange oil in a 64% yield.

¹H NMR (CDCl₃, 300 MHz) δ 2.9 (m, 1H), 2.6-2.1 (m, 20H), 1.61 (m, 2H),1.4 (m, 4H), 1.2 (m, 2H).

FI-MS (APCI): m/z observed in a positive mode: 268.2.

Intermediate 19 (±)-3-Nitro-9-(oxiran-2-ylmethyl)carbazole

Protocol B (using DEAD, PPh₃):

At rt under an inert atmosphere, (±)-glycyclol (0.018 g, 20 uL, 0.24mmol) is added into a solution of 3-nitro-9H-carbazole (0.050 g, 0.24mmol) in anhydrous THF (5 mL). Then PPh₃ (0.126 g, 0.48 mmol, 2.0equiv.) is added neat. The resulting mixture is allowed to cool to 0° C.(ice bath) and DEAD (0.084 g, 0.48 mmol, 2.0 equiv.) is added. Theresulting reaction mixture is allowed to stir at rt for 16 hours. Thesolvent is evaporated and the crude compound is purified via flashchromatography using Biotage device and EtOAc/cyclohexane (1/3) aseluant to give the title compound in a 95% yield as a yellow solid. ¹HNMR (DMSO d-₆, 300 MHz) δ 9.20 (d, 1H, J=2.26 Hz), 8.47-8.40 (m, 1H),8.34 (dd, 1H, J=9.04, 2.26 Hz), 7.93-7.74 (m, 2H), 7.65-7.53 (m, 1H),4.96 (dd, 1H, J_(AB)=15.64 Hz, J_(AX)=2.83 Hz), 4.54 (dd, 1H,J_(AB)=15.64 Hz, J_(AX)=5.84 Hz), 3.43-3.35 (m, 1H), 2.79 (t, 1H, J=4.52Hz), 2.58 (dd, 1H, J=4.9, 2.64 Hz).

Protocol B (using CMMP):

The same method as employed in the preparation of Intermediate 15 butstarting from 3-nitro-9H-carbazole gives the title compound in a 50%yield as a yellow powder. Heating up the reaction mixture in THF up to80° C. leads to the formation of3-nitro-9-[(1E)-3-(3-nitrocarabazol-9-yl)propen-1-enyl]carbazole in a10% yield. Using glycyclol (1 equiv.), CMMP (2 equiv.) and3-nitro-9H-carbazole (2 equiv.) in toluene at 110° C. gives3-nitro-9-[(1E)-3-(3-nitrocarabazol-9-yl)propen-1-enyl]carbazole in a90% yield.

-   3-Nitro-9-[(1E)-3-(3-nitrocarbazol-9-yl)propen-1-enyl]carbazole:

¹H NMR (DMSO d-₆, 300 MHz) δ 9.26 (d, 1H, J=2.26 Hz), 9.15 (d, 1H,J=2.07 Hz), 8.5-8.24 (m, 4H), 8.09 (d, 1H, J=9.03 Hz), 8.04 (d, 1H,J=8.28 Hz), 7.99-7.74 (m, 3H), 7.70-7.45 (m, 2H), 7.44-7.2 (m, 2H),6.5-6.35 (dt, 1H, J=13.94 Hz, 6.92 Hz), 5.47 (d, 2H, J=6.6 Hz)

FT-MS (APCI) m/z observed in a negative mode: 461.0.

Example 1(±)-1-(4-Benzyl-piperazin-1-yl)-3-(2-methyl-carbazol-9-yl)-propan-2-ol

To a solution of 2-methyl-9H-carbazole (0.5 g, 2.76 mmol) in 10 mL ofTHF is added n-BuLi (1.6 M in hexanes, 1.8 mL, 1.1 eq) under inertatmosphere. After 15 min of stirring at rt a solution of Intermediate 1in 4 mL of THF is added. After 16 hr at rt the reaction is judged to becompleted by tlc monitoring (SiO₂, cHex:EtOAc 80:20) and is quenchedwith 10 mL of a saturated aqueous solution of K₂CO₃. The reactionmixture is extracted with DCM, washed with brine (10 mL), dried overMgSO₄ and concentrated in vacuo. Flash chromatography on a 2×20 cm²column using cHex:EtOAc (80:20) as eluting solvent and removal of thesolvent in vacuo gives the title compound as a foam. Slow addition ofHCl (10 mL, 1M in Et₂O) into a solution of the above compound in EtOH (3mL) gives the hydrochloride salt of the title compound (0.39 g) as abeige powder in a 35% yield.

Mp: 239° C. (decomposition).

Analysis for C₂₇H₃₁N₃O. 2HCl:

Calculated: C, 66.66; H, 6.84; N, 8.64;

Found: C, 66.45; H, 6.92; N, 8.51%:

Example 2(±)-4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazine-1-carboxylictert-butyl ester

The same method as employed in the preparation of Example 1 but startingfrom 9H-carbazole and (±)-4-oxiranylmethyl-piperazine-1-carboxylic acidtert-butyl ester (prepared according to the procedure of Toldy, L. etal., WO 97/14685) gives after flash chromatography the title compound asa pale yellow foam in a 82% yield.

¹H NMR (CDCl₃, 300 MHz) δ 8.07 (d, 2H, J=7.8 Hz), 7.45 (m, 4H), 7.20 (m,2H), 4.37 (d, 2H, J=5.3 Hz), 4.20 (m, 1H), 3.38 (br s, 5H), 2.54-2.44(m, 4H), 2.40 (m, 2H), 1.44 (s, 9H).

Example 3 (±)-1-Carbazol-9-yl-3-piperazin-1-yl-propan-2-ol

At rt to a solution of Example 2 (3.4 g, 8.30 mmol) in DCM (120 mL) isadded TFA (30 mL). The resulting mixture is stirred at rt for 30 min.Concentration in vacuo gives an oily residue. Flash chromatography on a7×18 cm² column of SiO₂ using DCM:MeOH:TEA (40:5:3) as eluting solventand removal of the solvent gives the title compound (3.12 g, 92%) as awhite foam. Slow addition of HCl (30 mL, 4 eq, 1M in Et₂O) into asolution of the above compound in EtOH (40 mL) gives the hydrochloridesalt of the title compound as beige crystals in a quantitative yield.

M.p.: 245-250° C. (decomposition).

Analysis for C₁₉H₂₃N₃O. 2HCl. 0.65H₂O:

Calculated: C, 57.91; H, 6.73; N, 10.60; Cl, 17.99;

Found: C, 57.93; H, 6.67; N, 10.61; Cl, 18.03%.

Example 4(±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole,(±)-4-oxiranylmethyl-piperazine-1-carboxylic acid tert-butyl ester andsodium hydride gives after flash chromatography the title compound as awhite foam in a 72% yield. M.p: 90-95° C.

¹H NMR (CDCl₃, 300 MHz) δ 8.08 (d, 2H, J=1.9 Hz), 7.51 (dd, 2H, J=8.7,1.9 Hz), 7.32 (d, 2H, J=8.7 Hz), 4.3-4.1 (m, 3H), 3.36 (br s, 4H), 2.50(m, 2H), 2.36 (m, 2H), 2.26 (m, 2H), 1.45 (s, 9H).

Example 5 (±)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3,6-dibromocarbazol-9-yl-2-hydroxy-propyl)-piperazine-1-carboxylictert-butyl ester gives after flash chromatography the trifluoro acetatesalt of the title compound as a white solid in a 63% yield.

M.p.: 133° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 8.84 (br s, 2H), 8.46 (d, 2H, J=1.7 Hz),7.67-7.58 (m, 4H), 4.43-4.30 (m, 2H), 4.18 (br s, 1H), 3.18 (br s, 4H),2.93 (br s, 6H).

Analysis for C₁₉H₂₁Br₂N₃O. 2CF₃CO₂H. 0.13Et₂O:

Calculated: C, 40.05; H, 3.46; N, 5.97;

Found: C, 40.43; H, 3.85; N, 6.33%.

Example 6(±)-1-(4-Benzyl-piperazin-1-yl)-3-(3,6-dibromo-carbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole and sodium hydride gives after flashchromatography the title compound as a white foam in a 66% yield. M.p.:70-75° C.

Analysis for C₂₆H₂₇Br₂N₃O:

Calculated: C, 56.03; H, 4.88; N, 7.54;

Found: C, 55.81; H, 4.97; N, 7.36%.

Example 7 (±)-1-(4-Benzyl-piperazin-1-yl)-3-carbazol-9-yl-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 9H-carbazole gives the title compound as white crystals in a 66%yield.

M.p.: 205-210° C. (decomposition).

Analysis for C₂₆H₂₉N₃O. 2HCl. 0.4H₂O. 0.13EtOAc:

Calculated: C, 64.86; H, 6.74; N, 8.56; Cl, 14.44;

Found: C, 64.74; H, 6.83; N, 8.42; Cl, 14.54%.

Example 8(±)-4-[3-(3-Bromo-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester

The same method as employed in the preparation of Example 1 but startingfrom 3-bromo-9H-carbazole (prepared according to the procedure of Smith,K. et al Tetrahedron 1992, 48(36); pp 7474-7488),(±)-4-oxiranylmethyl-piperazine-1-carboxylic acid tert-butyl ester andsodium hydride gives after flash chromatography the title compound as apale yellow foam in a 62% yield. M.p.: 65-75° C.

¹H NMR (CDCl₃, 300 MHz) δ 8.16 (d, 1H, J=1.9 Hz), 7.99 (d, 1H, J=7.8Hz), 7.52 (dd, 1H, J=8.7, 1.9 Hz), 7.47 (m, 1H), 7.36 (d, 1H, J=8.7 Hz),7.25 (m, 2H), 4.33 (m, 2H), 4.20 (m, 1H), 3.38 (br s, 5H), 2.52 (m, 2H),2.46 (d, 2H, J=6.7 Hz), 2.32 (m, 2H), 1.44 (s, 9H).

Example 9 (±)-1-(3-Bromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3-bromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 63%yield. M.p.: 230-235° C. (decomposition).

¹H NMR (DMSO-d6, 300 MHz) δ 8.39 (d, 1H, J=1.8 Hz), 8.18 (d, 1H, J=7.8Hz), 7.70 (m, 2H), 7.50 (dd, 1H, J=8.7, 1.8 Hz), 7.45 (t, 1H, J=7.5 Hz),7.22 (t, 1H, J=7.4 Hz), 4.40 (br s, 3H), 3.38 (br s, 10H).

Example 10(±)-1-(4-Benzyl-piperazin-1-yl)-3-(2-hydroxy-carbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 9H-carbazol-2-ol and 2 eq of n-BuLi (according to the procedure ofAlbanese, D. et al Tetrahedron 1995, 51(19), pp 5681-5688) gives afterflash chromatography and recrystallization from EtOH the title compoundas a beige solid in a 30% yield.

M.p.: 202° C.

¹H NMR (DMSO-d6, 300 MHz) δ 9.42 (s, 1H), 7.91 (d, 1H, J=7.5 Hz), 7.84(d, 1H, J=8.4 Hz), 7.48 (d, 1H, J=8.2 Hz), 7.34-7.20 (m, 6H), 7.08 (t,1H, J=7.1 Hz), 6.89 (d, 1H, J=1.9 Hz), 6.65 (dd, 1H, J=8.3, 2.0 Hz),4.89 (d, 1H, J=4.9 Hz), 4.32 (dd, 1H, J=14.4, 3.6 Hz), 4.15-4.0 (m, 2H),3.31 (s, 2H), 3.02-2.34 (m, 10H).

Example 11(S)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole, Intermediate 2 and sodium hydride givesthe title compound as a white foam in a 61% yield. M.p.: 90-100° C.

¹H NMR (CDCl₃, 300 MHz) δ 8.11 (d, 2H, J=1.9 Hz), 7.54 (dd, 2H, J=8.7,1.9 Hz), 7.33 (d, 2H, J=8.7 Hz), 4.38-4.22 (m, 3H), 3.41 (br s, 4H),2.54 (br s, 2H), 2.43-2.33 (m, 4H), 1.41 (s, 9H).

96% ee (ChiralPak AD column, rt, isohexane:EtOH:TEA (90:10:0.1), 240 nM,1 mL/min).

Example 12(R)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole, Intermediate 3 and sodium hydride givesthe title compound as a white foam in a 62% yield. M.p.: 90-100° C.

¹H NMR (CDCl₃, 300 MHz) δ 8.11 (d, 2H, J=1.9 Hz), 7.54 (dd, 2H, J=8.7,1.9 Hz), 7.33 (d, 2H, J=8.7 Hz), 4.38-4.22 (m, 3H), 3.41 (br s, 4H),2.54 (br s, 2H), 2.43-2.33 (m, 4H), 1.41 (s, 9H).

98% ee (ChiralPak AD column, it, isohexane:EtOH:TEA (90:10:0.1), 240 nM,1 mL/min).

Example 13(S)-4-[3-(3-Bromo-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester

The same method as employed in the preparation of Example 1 but startingfrom 3-bromo-9H-carbazole, Intermediate 2 and sodium hydride gives thetitle compound as a white foam in a 61% yield. M.p.: 75-85° C.

¹H NMR (CDCl₃, 300 MHz) δ 8.16 (d, 1H, J=1.8 Hz), 8.02 (d, 1H, J=7.0Hz), 7.52 (dd, 1H, J=8.7, 1.9 Hz), 7.46 (m, 2H), 7.44 (d, 1H, J=8.7 Hz),7.3 (m, 1H), 4.39-4.24 (m, 3H), 3.41 (br s, 4H), 2.54 (br s, 2H),2.47-2.34 (m, 4H), 1.41 (s, 9H).

96% ee (ChiralPak AD column, rt, isohexane:EtOH:TEA (90:10:0.1), 240 nM,1 mL/min).

Example 14(R)-4-[3-(3-Bromo-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester

The same method as employed in the preparation of Example 1 but startingfrom 3-bromo-9H-carbazole, Intermediate 3 and sodium hydride gives thetitle compound as a white foam in a 64% yield. M.p.: 75-85° C.

¹H NMR (CDCl₃, 300 MHz) δ 8.16 (d, 1H, J=1.8 Hz), 8.02 (d, 1H, J=7.0Hz), 7.52 (dd, 1H, J=8.7, 1.9 Hz), 7.46 (m, 2H), 7.44 (d, 1H, J=8.7 Hz),7.3 (m, 1H), 4.39-4.24 (m, 3H), 3.41 (br s, 4H), 2.54 (br s, 2H),2.47-2.34 (m, 4H), 1.41 (s, 9H).

99% ee (ChiralPak AD column, rt, isohexane:EtOH:TEA (90:10:0.1), 240 nM,1 mL/min).

Example 15 (S)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(S)-4-[3-(3,6-dibromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 98%yield. M.p.: 304-305° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 11.0 (br s, 1H), 9.56 (br s, 2H), 8.46 (d,2H, J=1.9 Hz), 7.72 (d, 2H, J=8.8 Hz), 7.60 (dd, 2H, J=8.8, 1.9 Hz),5.92 (br s, 1H), 4.40 (br s, 3H), 3.56-3.37 (m, 10H).

96% ee (ChiralPak AD column, rt, isohexane:EtOH:TEA (90:10:0.1), 240 nM,1 mL/min).

Example 16 (R)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(R)-4-[3-(3,6-dibromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 99%yield. M.p.: 303-304° C.

¹H NMR (DMSO-d6, 300 MHz) δ 11.0 (br s, 1H), 9.56 (br s, 2H), 8.46 (d,2H, J=1.9 Hz), 7.72 (d, 2H, J=8.8 Hz), 7.60 (dd, 2H, J=8.8, 1.9 Hz),5.92 (br s, 1H), 4.40 (br s, 3H), 3.56-3.37 (m, 10H).

98% ee (ChiralPak AD column, rt, isohexane:EtOH:TEA (90:10:0.1), 240 nM,1 mL/min).

Example 17 (S)-1-(3-Bromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(S)-4-[3-(3-bromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 97%yield. M.p.: 77° C. (decomposition).

¹H NMR (DMSO-d₆, 300 MHz) δ 10.96 (br s, 1H), 9.59 (br s, 2H), 8.39 (d,1H, J=1.7 Hz), 8.20 (d, 1H, J=7.7 Hz), 7.70 (m, 2H), 7.57 (dd, 1H,J=8.7, 1.7 Hz), 7.47 (t, 1H, J=7.4 Hz), 7.22 (t, 1H, J=7.3 Hz), 5.91 (brs, 1H), 4.41 (br s, 3H), 3.54-3.36 (m, 10H).

96% ee (ChiralPak AD column, rt, isohexane:EtOH:TEA (80:20:0.1), 266 nM,1 mL/min).

Example 18 (R)-1-(3-Bromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(R)-4-[3-(3-bromocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 97%yield. M.p.: 70-74° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 10.96 (br s, 1H), 9.59 (br s, 2H), 8.39 (d,1H, J=1.7 Hz), 8.20 (d, 1H, J=7.7 Hz), 7.70 (m, 2H), 7.57 (dd, 1H,J=8.7, 1.7 Hz), 7.47 (t, 1H, J=7.4 Hz), 7.22 (t, 1H, J=7.3 Hz), 5.91 (brs, 1H), 4.41 (br s, 3H), 3.54-3.36 (m, 10H).

98% ee (ChiralPak AD column, it, isohexane:EtOH:TEA (80:20:0.1), 266 nM,1 mL/min).

Example 19(±)-1-[(3,4-Dichloro-phenyl)-piperazin-1-yl]-3-(2-methyl-carbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom Intermediate 4 gives the title compound as a pale pink powder in a20% yield.

M.p.: 140° C.

Analysis for C₂₆H₂₇N₃OCl₂. 2HCl. 0.9H₂O:

Calculated: C, 56.01; H, 5.57; N, 7.54;

Found: C, 56.02; H, 5.35; N, 7.67%.

Example 20(±)-1-[(3,4-Dichloro-phenyl)-piperazin-1-yl]-3-(carbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 9H-carbazole and Intermediate 4 gives the title compound as whitecrystals in a 26% yield. M.p.: 244° C. (decomposition).

Analysis for C₂₅H₂₅N₃OCl₂. HCl. 1.45H₂O. 0.16DCM:

Calculated: C, 56.96; H, 5.55; N, 7.92;

Found: C, 56.60; H, 5.12; N, 7.89%.

Example 21(±)-1-(4-Benzo[1,3]dioxol-5ylmethyl-piperazin-1-yl)-4-carbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 9H-carbazole and Intermediate 5 gives the title compound as a beigesolid in a 27% yield. M.p.: 170° C. (decomposition).

Analysis for C₂₇H₂₉N₃O₃. 1.9HCl. 0.7H₂O:

Calculated: C, 61.51; H, 6.21; N, 7.97;

Found: C, 61.38; H, 6.50; N, 7.88%.

Example 22(±)-1-Carbazol-9-yl-3-[4-(4-fluoro-benzyl)-piperazin-1-yl]-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 9H-carbazole and Intermediate 6 gives the title compound as a beigesolid in a 35% yield. M.p.: 251° C. (decomposition).

Analysis for C₂₆H₂₈FN₃O. 2HCl. H₂O:

Calculated: C, 61.42; H, 6.34; N, 8.26;

Found: C, 61.21; H, 6.42; N, 8.18%.

Example 23(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(4-fluoro-benzyl)-piperazin-1-yl]-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole, Intermediate 6 and sodium hydride givesthe title compound as white crystals in a 60% yield. M.p.: 297° C.(decomposition).

Analysis for C₂₆H₂₆Br₂FN₃O. 2HCl:

Calculated: C, 48.17; H, 4.35; N, 6.48;

Found: C, 47.91; H, 4.55; N, 6.21%.

Example 24(±)-1-[4-(4-Fluoro-benzyl)-piperazin-1-yl]-3-(3-phenyl-carbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but starting9H-carbazole, Intermediate 7 and Intermediate 6 gives the title compoundas white crystals in a 40% yield. M.p.: 188° C.

Analysis for C₃₂H₃₂FN₃O. 2HCl. 2H₂O:

Calculated: C, 63.78; H, 6.36; N, 6.97;

Found: C, 63.76; H, 6.23; N, 7.02%.

Example 25(±)-9-(2-Hydroxy-3-piperazin-1-yl-propyl)-carbazole-3,6-dicarbonitrile

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3,6-dicyanocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 63%yield. M.p.: 295° C. (decomposition).

Analysis for C₂₁H₂₁N₅O. 2HCl. 1.8H₂O:

Calculated: C, 54.27; H, 5.77; N, 15.07;

Found: C, 54.25; H, 6.08; N, 14.98%.

4-[3-(3,6-Dicyanocarbazol-9-yl)-2-hydroxy-propyl]piperazine-1-carboxylictert-butyl ester (0.38 g, 52%) is obtained as a yellow solid using thesame method as employed in the preparation of Example 1 but startingfrom 4-oxiranylmethyl-piperazine-1-carboxylic acid tert-butyl ester(0.20 g, 0.83 mmol), 9H-carbazole-3,6-dicarbonitrile (0.20 g, 0.92 mmol)(prepared according to the procedure of Patrick, D. A. et al Eur. J.Med. Chem. 1997, 32, pp 781-793) and sodium hydride (40 mg, 0.92 mmol).

¹H NMR (CDCl₃, 300 MHz) δ 8.33 (d, 2H, J=1.3 Hz), 7.71 (dd, 2H, J=8.6,1.3 Hz), 7.30 (d, 2H, J=8.6 Hz), 4.42 (dd, 1H, J=15.1, 3.1 Hz), 4.34(dd, 1H, J=15.1, 6.3 Hz), 4.18 (br s, 1H), 3.56 (br s, 1H), 3.37 (m,4H), 2.56-2.36 (m, 6H), 1.44 (s, 9H).

Example 26 (±)-1-(3-Nitrocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3-nitrocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a yellow solid in a 95%yield. M.p.: 112° C. (decomposition).

¹H NMR (DMSO-d₆:D₂O (700:40), 300 MHz) δ 9.17 (d, 1H, J=2.1 Hz), 8.40(d, 1H, J=7.7 Hz), 8.34 (dd, 1H, J=9.1, 2.1 Hz), 7.84 (d, 1H, J=9.1 Hz),7.80 (d, 1H, J=8.3 Hz), 7.57 (t, 1H, J=7.6 Hz), 7.33 (t, 1H, J=7.5 Hz),4.50 (m, 2H), 4.36 (br s, 1H), 3.47-3.08 (m, 10H).

(±)-4-[3-(3-Nitrocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester (0.135 g, 43%) is obtained as a yellow foam using thesame method as employed in the preparation of Example 1 but startingfrom 3-nitro-9H-carbazole (0.218 g, 1.03 mmol) (prepared according tothe procedure of Kyziol, J. B. et al Tetrahedron 1984, 40(10), pp1857-1861), (±)-4-oxiranylmethyl-piperazine-1-carboxylic acid tert-butylester (0.166 g, 0.685 mmol) and sodium hydride (0.045 g, 1.027 mmol).

¹H NMR (CDCl₃, 300 MHz) δ 8.93 (d, 1H, J=3.1 Hz), 8.34 (dd, 1H, J=9.1,2.7 Hz), 8.10 (d, 1H, J=7.9), 7.53-7.49 (m, 3H), 7.30 (ddd, 1H, J=7.8,6.6, 1.6 Hz), 4.43 (dd, 1H, J=15.1, 4.3 Hz), 4.34 (dd, 1H, J=15.1, 5.8Hz), 4.20 (m, 1H), 3.37 (m, 4H), 2.54 (m, 2H), 2.37 (m, 2H), 2.30 (m,2H), 1.44 (s, 9H).

Alternatively, the same method as employed in the preparation of Example73 could be employed, but starting from Intermediate 19 and piperazinein EtOH gives the hydrochloride of the title compound as a yellowcompound in a one step procedure in 98% yield.

M.p.: 112° C.

¹H NMR (DMSO-d6, 300 MHz) δ 9.17 (d, 1H, J=2.1 Hz), 8.40 (d, 1H, J=7.7Hz), 8.34 (dd, 1H, J=9.1, 2.1 Hz), 7.84 (d, 1H, J=9.1 Hz), 7.80 (d, 1H,J=8.3 Hz), 7.57 (t, 1H, J=7.6 Hz), 7.33 (t, 1H, J=7.5 Hz), 4.50 (m, 2H),4.36 (br s, 1H), 3.47-3.08 (m, 10H).

Example 27 (±)-1-(3-Phenylcarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3-phenylcarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 78%yield. M.p.: 120° C.

¹H NMR (DMSO-d₆:D₂O (700:40), 300 MHz) δ 8.46 (s, 1H), 8.24 (d, 1H,J=7.7 Hz), 7.78-7.76 (m, 4H), 7.70 (d, 1H, J=8.2 Hz), 7.48 (m, 3H), 7.32(t, 1H, J=7.3 Hz), 7.22 (t, 1H, J=7.5 Hz), 4.40 (m, 3H), 3.42-3.31 (m,10H).

(±)-4-[3-(3-Phenylcarbazol-9-yl)-2-hydroxy-propyl]piperazine-1-carboxylictert-butyl ester (0.135 g, 35%) is obtained as a white foam using thesame method as employed in the preparation of Example 1 but startingfrom Intermediate 7 (0.182 g, 0.75 mmol) and(±)-4-oxiranylmethyl-piperazine-1-carboxylic acid tert-butyl ester(0.165 g, 0.68 mmol).

¹H NMR (CDCl₃, 300 MHz) δ 8.30 (d, 1H, J=1.7 Hz), 8.12 (d, 1H, J=7.7Hz), 7.71 (m, 3H), 7.53-726 (m, 7H), 4.38 (m, 2H), 4.22 (m, 1H), 3.52(br s, 4H), 2.54-2.42 (m, 4H), 2.30 (m, 2H), 1.44 (s, 9H).

Example 28 (±)-1-(2-Hydroxycarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(2-hydroxycarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a beige solid in a 10%yield.

¹H NMR (DMSO-d₆, 300 MHz) δ 9.50 (br s, 1H), 7.65 (d, 1H, J=7.4 Hz),7.86 (d, 1H, J=8.3 Hz), 7.55 (d, 1H, J=8.2 Hz), 7.30 (t, 1H, J=7.2 Hz),7.12 (t, 1H, J=7.4 Hz), 6.96 (s, 1H), 6.68 (dd, 1H, J=8.3, 1.6 Hz), 4.27(br s, 3H), 2.49 (m, 10H).

(±)-4-[3-(2-Hydroxycarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester is obtained as an oil which is used withoutpurification using the same method as employed in the preparation ofExample 1 but starting from 2-hydroxy-9H-carbazole (0.182 g, 0.75 mmol),2 eq of n-BuLi (2.2 mL, 1.6 M in Hexanes) (according to the procedure ofAlbanese, D. et al Tetrahedron 1995, 51(19), pp 5681-5688) and4-oxiranyl-methyl-pipera-zine-1-carboxylic acid tert-butyl ester (0.210g, 0.87 mmol).

Example 29(±)-1-(3,6-Diphenylcarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom4-[3-(3,6-diphenylcarbazol-9-yl-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 88%yield.

M.p.: 270-288° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 8.39 (s, 2H), 7.56 (m, 8H), 7.27 (m, 4H),7.11 (t, 2H, J=7.3 Hz), 4.25 (m, 3H), 3.30 (m, 10H).

4-[3-(3,6-Diphenylcarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester (0.110 g, 60%) is obtained as a white solid using thesame method as employed in the preparation of Example 1 but startingfrom Intermediate 8 (0.114 g, 0.36 mmol),(±)-4-oxiranylmethyl-piperazine-1-carboxylic acid tert-butyl ester(0.079 g, 0.326 mmol) and sodium hydride (0.017 g, 0.39 mmol).

¹H NMR (CDCl₃, 300 MHz) δ 8.32 (d, 2H, J=1.2 Hz), 7.73-7.69 (m, 6H),7.54-7.27 (m, 8H), 4.43 (br s, 2H), 4.11 (s, 1H), 3.34 (br s, 4H), 2.45(br s, 4H), 2.02 (br s, 2H), 1.44 (s, 9H).

Example 30(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(3-phenyl-propyl)-piperazin-1-yl]-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole, Intermediate 9 and sodium hydride givesthe title compound as a white solid in a 35% yield. M.p.: 255° C.

Analysis for C₂₈H₃₁BrN₃O. 2HCl. 0.7H₂O:

Calculated: C, 50.13; H, 5.17; N, 6.26;

Found: C, 50.09; H, 5.54; N, 5.92%.

Example 31(±)-1-Carbazol-9-yl-3-[4-(3-phenyl-propyl)-piperazin-1-yl]-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 9H-carbazole, Intermediate 9 and sodium hydride gives the titlecompound as a white solid in a 35% yield. M.p.: 265° C.

Analysis for C₂₈H₃₃N₃O. 2HCl. 0.5H₂O:

Calculated: C, 66.01; H, 7.12; N, 8.25;

Found: C, 66.11; H, 7.26; N, 8.27%.

Example 32(±)-3,6-Dibromo-9-(2-fluoro-3-piperazin-1-yl-propyl)-carbazole

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3,6-dibromo-carbazol-9-yl)-2-fluoro-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 61%yield. M.p.: 270° C.

Analysis for C₁₉H₂₀Br₂FN₃. 2HCl. 0.5H₂O:

Calculated: C, 41.41; H, 4.21; N, 7.62;

Found: C, 41.29; H, 4.17; N, 7.51%.

(±)-4-[3-(3,6-Dibromo-carbazol-9-yl)-2-fluoro-propyl]-piperazine-1-carboxylictert-butyl ester (0.05 g, 80%) is obtained as a colorless oil from thereaction of Example 4 (0.100 g, 0.176 mmol) and DAST (100 μl., 0.763mmol) in DCM (4 mL) at 0° C. Quenching with a saturated aqueous solutionof NaHCO₃, extraction with DCM, drying over MgSO₄, flash chromatographyon a 2×15 cm² column of SiO₂ using DCM:MeOH (100:1.5) as eluting solventand concentration in vacuo gives the above compound.

¹H NMR (CDCl₃, 300 MHz) δ 8.05 (d, 2H, J=1.9 Hz), 7.50 (dd, 2H, J=8.7,1.9 Hz), 7.29 (d, 2H, J=8.7 Hz), 4.93 (dm, 1H, J=47.3 Hz), 4.85 (m,0.5H), 4.53-4.46 (m, 2H), 3.45 (m, 4H), 2.54 (m, 2H), 2.41 (br s, 4H),1.44 (s, 9H).

Example 33 (±)-1-(3-Amino-carbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3-amino-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives after the title compound as a beige solid in a61% yield. M.p.: 220° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 10.49 (br s, 2H), 9.79 (br s, 1H), 8.18 (d,1H, J=7.7 Hz), 8.11 (d, 1H, J=1.7 Hz), 7.83 (d, 1H, J=8.7 Hz), 7.75 (d,1H, J=8.3 Hz), 7.50 (m, 2H), 7.27 (m, 2H), 7.06 (s, 1H), 5.88 (br s,1H), 4.44 (br s, 3H), 3.49-3.15 (m, 10H).

(±)-4-[3-(3-Amino-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester is obtained from the reduction of(±)-4-[3-(3-nitrocarbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester (0.100 g, 0.22 mmol) using SnCl₂.H₂O (0.248 g, 1.10mmol) in EtOH (2 mL) at 70° C. for 2 hours. Addition of an aqueoussaturated solution of NaHCO₃ until pH=7-8, extraction with EtOAc, dryingover MgSO₄ and concentration in vacuo gives a brown oil. Purificationvia flash chromatography on a 2×15 cm² column (SiO₂, DCM:MeOH (97:3))gives the above compound as an orange oil in a 58% yield.

¹H NMR (CDCl₃, 300 MHz) δ 7.97 (d, 1H, J=7.7 Hz), 7.42-7.38 (m, 3H),7.27 (d, 1H, J=8.5 Hz), 7.15 (m, 1H), 6.88 (dd, 1H, J=8.5, 2.2 Hz), 4.31(m, 2H), 4.20 (m, 1H), 3.37 (m, 4H), 2.55-2.40 (m, 4H), 2.27 (m, 2H),1.41 (s, 9H).

Example 34(±)-N-[9-(2-Hydroxy-3-piperazin-1-yl-propyl)-carbazol-3-yl]-acetamide

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3-acetylamino-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a pale grey powder in a 63%yield. M.p.>180° C. (decomposition).

¹H NMR (DMSO-d₆:D₂O (700:40), 300 MHz) δ 8.34 (d, 1H, J=1.8 Hz), 8.03(d, 1H, J=7.7 Hz), 7.66 (d, 1H, J=8.3 Hz), 7.62 (d, 1H, J=8.8 Hz), 7.55(dd, 1H, J=8.8, 1.8 Hz), 7.39 (t, 1H, J=7.5 Hz), 7.17 (t, 1H, J=7.5 Hz),4.49 (m, 1H), 4.37 (m, 2H), 3.64-3.28 (m, 10H), 2.06 (s, 3H).

(±)-4-[3-(3-Acetylamino-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylictert-butyl ester is obtained from the reaction of(±)-4-[3-(3-amino-carbazol-9-yl)-2-hydroxy-propyl]-piperazine-1-carboxylicacid tert-butyl ester (0.139 g, 0.33 mmol) and acetic acid (21 μL, 0.33mmol) in the presence of HOBt (0.056 g, 0.36 mmol), EDCI.HCl (0.069 g,0.36 mmol), TEA (50 μL, 0.33 mmol) in DCM. Standard quenching andwork-up gives the above compound as a brown foam.

¹H NMR (CDCl₃, 300 MHz) δ 8.16 (d, 1H, J=1.9 Hz), 7.90 (m, 2H), 7.37 (m,3H), 7.27 (d, 1H, J=8.7 Hz), 7.13 (m, 1H), 4.30-4.06 (m, 3H), 3.32 (m,4H), 2.43 (m, 2H), 2.34 (m, 2H), 2.22 (m, 2H), 2.13 (s, 3H), 1.42 (s,9H).

Example 35(±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-2-phenoxy-ethanone

To a solution of Example 3 (0.200 g, 0.6 mmol) in 25 mL of DCM are addedTEA (90 μL, 1 eq), EDCI (0.124 g, 1 eq), HOBT (0.087 g, 1 eq) andphenoxy-acetic acid (0.099 g, 1 eq). After 12 hr of stirring at rt thereaction is judged to be complete by tlc monitoring (SiO₂, DCM:MeOH(95:5)) and is quenched with 10 mL of water. The reaction mixture isextracted with DCM, washed with brine (5 mL), dried over MgSO₄ andconcentrated in vacuo. Flash chromatography on a 2.5×12 cm² column ofsilica gel using DCM:MeOH (95:5) as eluting solvent and removal of thesolvent in vacuo gives the title compound as a white foam. Slow additionof HCl (5 mL, 1M in Et₂O) into a solution of the above compound in DCM(2 mL) gives the hydrochloride salt of the title compound (0.39 g) as abeige powder in a 66% yield.

M.p.: 130° C.

¹H NMR (CDCl₃, 300 MHz) δ 10.22 (br s, 1H), 8.14 (d, 2H, J=8.2 Hz), 7.68(d, 2H, J=8.2 Hz), 7.46 (tr, 2H, J=7.2 Hz), 7.42-7.18 (m, 4H), 6.95-6.89(m, 3H), 6.00 (br s, 1H), 4.83 (s, 2H), 4.4-3.9 (m, 5H), 3.7-3.1 (m,8H).

Example 36(±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-1-phenyl-methanone

The same method as employed in the preparation of Example 35 butstarting from benzoic acid gives the title compound as a white powder ina 56% yield. M.p.: 153° C.

Analysis for C₂₆H₂₇N₃O₂. HCl. 1.3H₂O:

Calculated: C, 65.97; H, 6.51; N, 8.88;

Found: C, 65.96; H, 6.35; N, 8.86%.

Example 37(±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-2-(4-hydroxy-phenoxy)-ethanone

The same method as employed in the preparation of Example 35 butstarting from 4-hydroxyphenoxy acetic acid gives the title compound as awhite powder in a 27% yield. M.p.: 155° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 10.25 (br s, 1H), 9.96 (s, 1H), 8.14 (d, 2H,J=7.7 Hz), 7.67 (d, 2H, J=8.2 Hz), 7.47-7.42 (m, 2H), 7.29-7.18 (m, 4H),6.80 (d, 2H, J=8.6 Hz), 6.01 (br s, 1H), 4.48 (m, 3H), 4.11 (br s, 2H),3.52-3.10 (m, 10H).

Example 38(±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-1-(4-hydroxy-phenyl)-methanone

The same method as employed in the preparation of Example 35 butstarting from 4-hydroxybenzoic acid gives the title compound as a whitepowder in a 68% yield.

M.p.: 184° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 9.96 (br s, 1H), 8.89 (s, 1H), 8.09 (d, 2H,J=7.7 Hz), 7.61 (d, 2H, J=8.2 Hz), 7.43-7.37 (m, 2H), 7.15 (m, 2H),6.69-6.57 (m, 4H), 5.95 (br s, 1H), 4.63 (s, 1H), 4.30 (br m, 3H),3.47-3.04 (m, 10H).

Example 39(±)-1-[4-(3-Carbazol-9-yl-2-hydroxy-propyl)-piperazin-1-yl]-1-(4-fluorophenyl)-methanone

The same method as employed in the preparation of Example 35 butstarting from 4-fluoro-benzoic acid gives the title compound as a whitepowder in an 86% yield.

M.p.: 70-80° C.

Analysis for C₂₆H₂₆N₃O₂. HCl. 0.5H₂O. 0.17Et₂O:

Calculated: C, 65.46; H, 6.11; N, 8.58;

Found: C, 65.27; H, 6.38; N, 8.41%.

Example 40(±)-1-(4-Benzenesulfonyl-piperazin-1-yl)-3-carbazol-9-yl-propan-2-61

To a solution of Example 3 (0.200 g, 0.6 mmol) in 20 mL of DMF wereadded TEA (100 μL, 1 eq) and benzene sulfonyl chloride (0.114 g, 1 eq).After 12 hr of stirring at rt the reaction is judged to be complete bytlc monitoring (SiO₂, DCM:MeOH (95:5)) and is concentrated in vacuo. Theresidue is taken up in DCM (10 mL), washed with brine (5 mL) andextracted. The combined organic layers were dried over MgSO₄ andconcentrated in vacuo. Flash chromatography on a 2.5×12 cm² column ofsilica gel using DCM:MeOH (95:5) as eluting solvent and removal of thesolvent in vacuo gives the title compound as a white powder. Slowaddition of HCl (4 eq, 1M in Et₂O) into a solution of the above compoundin DCM (2 mL) gives the hydrochloride salt of the title compound (0.39g) as white crystals in a 92% yield. M.p.: 130° C.

Analysis for C₂₅H₂₇N₃O₃S. HCl. 0.5H₂O:

Calculated: C, 60.66; H, 5.90; N, 8.49;

Found: C, 60.53; H, 5.94; N, 8.34%.

Example 41(±)-9-[3-(4-Benzyl-piperazin-1-yl)-2-methoxy-propyl]-carbazole

To a solution of Example 7 (0.200 g, 0.50 mmol) in 5 mL of THF are addedNaH (0.068 g, 1.56 mmol) and methyl iodide (125 μL, 2.0 mmol). After 2hr of stirring at rt the reaction is judged to be complete by tlcmonitoring (SiO₂, DCM:MeOH (100:3)) and is quenched with a saturatedaqueous solution of Na₂CO₃ (5 mL). Extraction with DCM, drying overMgSO₄, concentration in vacuo, flash chromatography on a 2.5×12 cm²column of silica gel using DCM:MeOH (95:5) as eluting solvent andremoval of the solvent in vacuo gives the title compound as an oil. Slowaddition of HCl (4 eq, 1M in Et₂O) into a solution of the above compoundin EtOH (2 mL) gives the hydrochloride salt of the title compound (0.39g) as white crystals in a 60% yield. M.p.: 272° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 8.14 (d, 2H, J=7.7 Hz), 7.67 (d, 2H, J=8.2Hz), 7.60 (br s, 2H), 7.46 (m, 5H), 7.20 (tr, 2H, J=7.4 Hz), 4.40 (br s,2H), 4.32 (br s, 3H), 3.75-3.10 (br m, 8H), 3.04 (s, 3H).

Example 42(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-piperazin-1-yl-propan-2-one

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3,6-dibromo-carbazol-9-yl)-2-oxo-propyl]-piperazine-1-carboxylictert-butyl ester gives the title compound as a white solid in a 83%yield. M.p.: 175° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 9.31 (br s, 2H), 8.48 (d, 2H, J=1.6 Hz),7.59 (dd, 2H, J=8.8, 1.6 Hz), 7.54 (d, 2H, J=8.8 Hz), 5.52 (s, 2H), 4.18(m, 2H), 3.25-3.13 (m, 8H).

(±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-oxo-propyl]-piperazine-1-carboxylictert-butyl ester (0.0.38 g, 38%) is obtained as a colorless oil by Swernoxidation of Example 4 (0.100 g, 0.176 mmol) using oxalyl chloride (20μL, 0.211 mmol), DMSO (31 μL, 0.44 mmol) and TEA (125 μL, 0.90 mmol) inDCM (1 mL) at −78° C.

¹H NMR (CDCl₃, 300 MHz) δ 8.09 (d, 2H, J=1.9 Hz), 7.51 (dd, 2H, J=8.7,1.9 Hz), 7.10 (d, 2H, J=8.7 Hz), 4.99 (s, 2H), 3.39 (m, 4H), 3.03 (s,2H), 2.30 (m, 4H), 1.43 (s, 9H).

Example 43(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(2-hydroxy-3-methylamino-propyl)-piperazin-1-yl]-propan-2-ol

To(±)-1-(3,6-dibromo-carbazol-9-yl)-3-(4-oxiranylmethyl-piperazin-1-yl)-propan-2-ol(0.110 g, 0.21 mmol) is added methyl amine (3 mL, 2M in MeOH). Theresulting mixture was stirred for 16 hr at rt. Concentration in vacuo,flash chromatography on a 2×14 cm² column of SiO₂ using (ACN:(aqueousNH₄OH-25%)) (5:1) as eluting solvent gives the title compound as a whitefoam in a 90% yield. M.p.: 65-75° C. Slow addition of HCl (1 mL, 1M inEt₂O) into a solution of the above compound in DCM (2 mL) gives thehydrochloride salt of the title compound (0.39 g) as a white solid in a35% yield.

M.p.>200° C.

¹H NMR (DMSO-d₆, 300 MHz) δ 8.78 (br s, 2H), 8.41 (d, 2H, J=1.8 Hz),7.66 (d, 2H, J=8.8 Hz), 7.54 (dd, 2H, J=8.8, 1.8 Hz), 4.36-2.83 (m,18H), 2.43 (s, 3H).

(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-(4-oxiranylmethyl-piperazin-1-yl)-propan-2-ol(0.118 g, 0.24 mmol) is obtained after chromatography on a 2×20 cm²column of SiO₂ using DCM:MeOH (97:3) as eluting solvent, as a whitesolid using the same method as employed in the preparation of Example 1but starting from 3,6-dibromo-9H-carbazole (0.30 g, 0.92 mmol),(±)-1,4-bis-oxiranylmethyl-piperazine (prepared according to theprocedure of Gerzon et al J. Med. Pharm. Chem. 1959, 1, p223) (0.20 g,1.01 mmol) and sodium hydride.

¹H NMR (CDCl₃, 300 MHz) δ 8.02 (d, 2H, J=1.9 Hz), 7.50 (dd, 2H, J=8.7,1.9 Hz), 7.25 (d, 2H, J=8.7 Hz), 4.22 (dd, 1H, J=14.9, 3.9 Hz), 4.13(dd, 1H, J=14.9, 5.9 Hz), 4.05 (m, 1H), 3.70 (d, 1H, J=11.6 Hz), 3.00(m, 1H), 2.71 (t, 1H, J=4.5 Hz), 2.65 (ddd, 1H, J=13.3, 7.4, 3.1 Hz),2.61-2.26 (m, 11H), 2.14 (ddd, 1H, J=13.3, 8.6, 7.0 Hz).

Example 44(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(3-phenyl-1,2,4-thiadiazol-5-yl)-piperazin-1-yl]-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole, Intermediate 10 and sodium hydride givesthe title compound as white crystals in a 28% yield. M.p.: 196° C.(decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.22 (d, 2H, J=1.8 Hz), 7.85 (m, 2H),7.44-7.36 (m, 5H), 7.23 (m, 2H), 4.17 (br s, 3H); 3.7 (br s, 2H), 3.38(br s, 8H).

Example 45(±)−1-4-Cyclohexylmethylpiperazin-1-yl)-3-(3,6-dibromo-carbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole, Intermediate 11 and sodium hydride andheating at 60° C. for 16 hours gives after flash chromatography thetitle compound as a white foam. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in DCM gives the hydrochloride salt ofthe title compound as a beige solid in a 35% yield.

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.45 (d, 2H, J=1.9 Hz), 7.68 (d, 2H,J=8.8 Hz), 7.60 (dd, 2H, J=8.7, 1.9 Hz), 4.4-4.3 (m, 3H), 3.6-3.0 (m,11H), 2.9 (br s, 1H), 1.8-1.5 (m, 6H), 1.2-0.9 (m, 5H).

Mp: 280° C. (decomposition).

Analysis for C₂₆H₃₃Br₂N₃O. 2HCl. 0.3 H₂O:

Calculated: C, 48.67; H, 5.69; N, 6.55;

Found: C, 48.46; H, 5.98; N, 6.42%:

Example 46(±)-1-[(4-Fluorophenyl)-piperazin-1-yl]-3-(3,6-dibromocarbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole, Intermediate 12 and sodium hydride andheating at 60° C. for 16 hours gives after flash chromatography thetitle compound as a yellow oil. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in DCM gives the hydrochloride salt ofthe title compound as a camel-colored solid in a 54% yield.

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.44 (d, 2H, J=1.7 Hz), 7.67-7.58 (m,4H), 7.10-6.95 (m, 4H), 4.6-4.2 (m, 5H), 3.8-3.5 (m, 5H), 3.4-3.0 (m,4H).

Mp: 230° C. (decomposition).

Analysis for C₂₅H₂₄Br₂FN₃O. 2HCl. 0.5H₂O:

Calculated: C, 46.68; H, 4.23; N, 6.53;

Found: C, 47.08; H, 4.67; N, 6.50%.

Example 47(±)-1-(Carbazol-9-yl)-3-[4-(2-fluorobenzyl)piperazin-1-yl]propan-2-ol

A solution of Example 3 (0.05 g, 0.16 mmol) in 25 mL of TMOF under anargon atmosphere is treated with 2-fluorobenzaldehyde (0.06 g, 0.48mmol) for 4 hours. Then sodium triacetoxy borohydride (0.103 g, 0.48mmol) is added neat. After 2 hours of stirring at rt, the resultingreaction mixture is quenched with water (10 mL). Extraction with 3×Et₂O(50 mL), drying over MgSO₄ and evaporation under reduced pressure givesan oily residue. Flash chromatography on a 2×20 cm² column of SiO₂ usingDCM/MeOH as eluting solvent (95/5) gives the title compound (0.08 g,0.19 mmol) as a colorless oil in a 60% yield. Slow addition of HCl, (4mL, 1M in Et₂O) into a solution of the above compound in MeOH (2 mL)gives the hydrochloride salt of the title compound.

Mp: 252° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.30-8.27 (d, 2H, J=7.7 Hz), 7.81-7.78(d, 2H, J=8.3 Hz), 7.64-7.54 (m, 4H), 7.41-7.33 (m, 4H), 4.53 (br s,3H), 4.03 (br s, 2H), 3.7-3.0 (m, 10H).

Example 48(±)-1-(Carbazol-9-yl)-3-[4-(4-nitrobenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from 4-nitrobenzaldehyde and Biotage purification gives an oilycompound. Slow addition of HCl (1M in Et₂O) into a solution of the abovecompound in MeOH gives the hydrochloride salt of the title compound(0.03 g, 0.07 mmol) as a yellow solid in a 20% yield.

Mp: 213° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.00 (d, 2H, J=8.6 Hz), 7.92 (d, 2H,J=7.8 Hz), 7.40 (tr, 4H, J=8.15, 7.0 Hz), 7.22 (doublet of tr, 2H,J=7.80, 1.0 Hz), 6.99 (tr, 2H, J=7.67, 7.38 Hz), 4.53 (br s, 3H), 3.60(br s, 2H), 3.4-2.9 (m, 10H).

Example 49(±)-1-(Carbazol-9-yl)-3-[4-(4-methoxybenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from 4-methoxybenzaldehyde and Biotage purification gives anoily compound. Slow addition of HCl (1M in Et₂O) into a solution of theabove compound in MeOH gives the hydrochloride salt of the titlecompound (0.02 g, 0.04 mmol) as a grey blue solid in a 20% yield.

Mp: 220° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 7.90 (d, 2H, J=8.6 Hz), 7.41 (d, 2H,J=8.25 Hz), 7.24-7.12 (m, 4H), 7.22 (tr, 2H, J=7.40 Hz), 6.78 (d, 2H,J=8.13 Hz), 4.20 (br s, 3H), 3.90 (br s, 2H), 3.4-2.9 (m, 10H).

Example 50(±)-1-[4-(4-Fluorobenzyl)piperazin-1-yl]-3-{3-[4-(trifluoromethyl)phenyl]-carbazol-9-yl}propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom Intermediate 6, Intermediate 13 and sodium hydride and heating at60° C. for 16 hours gives after Biotage chromatography the titlecompound as a yellow oil. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in DCM gives the hydrochloride salt ofthe title compound as a camel-colored solid in a 52% yield.

Mp: 237° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.54 (d, 1H, J=1.45 Hz), 8.22 (d, 1H,J=7.65 Hz), 7.97 (d, 2H, J=8.22 Hz), 7.82-7.71 (m, 4H), 7.64 (d, 1H,J=8.3 Hz), 7.46-7.42 (tr, 3H, J=7.2 Hz), 7.23-7.18 (tr, 4H, J=7.8 Hz),4.4-4.2 (m, 3H), 4.0 (m, 1H), 3.4-3.0 (m, 11H).

Example 51(±)-1-[4-(4-Fluorobenzyl)piperazin-1-yl]-3-{3-[4-(trifluoromethoxy)phenyl]-carbazol-9-yl}propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom Intermediate 6, Intermediate 14 and sodium hydride and heating at60° C. for 16 hours gives after Biotage chromatography the titlecompound as a yellow oil. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in DCM gives the hydrochloride salt ofthe title compound as a camel-colored solid in a 12% yield.

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.40 (d, 1H, J=1.28 Hz), 8.15 (d, 1H,J=7.82 Hz), 7.97 (d, 2H, J=8.76 Hz), 7.74-7.62 (m, 2H), 7.58 (d, 1H,J=8.3 Hz), 7.44-7.30 (m, 4H), 7.20-7.03 (m, 3H), 6.75 (d, 1H, J=8.4 Hz),4.4-4.2 (m, 3H), 4.0 (m, 1H), 3.4-3.0 (m, 11H).

Example 52(±)-1-(Carbazol-9-yl)-3-[4-(3-fluorobenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from 3-fluorobenzaldehyde using a parallel Quest 210synthesizer (0.32 mmol scale) and Biotage purifica-tion gives an oilycompound. Slow addition of HCl (1M in Et₂O) into a solution of the abovecompound in MeOH gives the hydrochloride salt of the title compound(0.128 g) as a beige-colored solid in a 95% yield.

Mp: 154° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.13 (d, 2H, J=7.6 Hz), 7.65 (m, 2H),7.46-7.18 (m, 8H), 4.63 (br s, 2H), 4.38 (br s, 1H), 3.42-3.12 (m, 12H).

Example 53(±)-1-(Carbazol-9-yl)-3-[4-(thien-2-ylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from thiophene-2-carboxaldehyde using a parallel Quest 210synthesizer (0.32 mmol scale) and Biotage purification gave an oilycompound. Slow addition of HCl (1M in Et₂O) into a solution of the abovecompound in MeOH gives the hydrochloride salt of the title compound(0.080 g) as a beige-colored solid in a 61% yield.

Mp: 248° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.12 (d, 2H, J=7.7 Hz), 7.63 (m, 3H),7.43 (tr, 2H, J=7.4, 7.9 Hz), 7.19 (tr, 3H, J=7.3 Hz), 7.05 (m, 1H),4.37 (br s, 2H), 4.25 (br s, 1H), 3.52-2.93 (m, 12H).

Example 54 (±)-1-(4-Butylpiperazin-1-yl)-(3-carbazol-9-yl)propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from butanal using a parallel Quest 210 synthesizer (0.32 mmolscale) and Biotage purification gives an oily compound. Slow addition ofHCl (1M in Et₂O) into a solution of the above compound in MeOH gives thehydrochloride salt of the title compound (0.017 g) as a white solid in a15% yield.

Mp: 242° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.12 (d, 2H, J=7.7 Hz), 7.62 (d, 2H,J=8.2 Hz), 7.42 (tr, 2H, J=7.21, 8.1 Hz), 7.19 (tr, 2H, J=7.4 Hz), 4.39(br s, 2H), 4.31 (br s, 1H), 3.52-3.01 (m, 12H), 1.6 (m, 2H), 1.34 (m,2H), 0.87 (tr, 3H, J=7.25 Hz).

Example 55(±)-4-({4-[3-carbazol-9-yl-2-hydroxypropyl]piperazin-1-yl}methyl)phenol

The same method as employed in the preparation of Example 47 butstarting from 4-hydroxybenzaldehyde using a parallel Quest 210synthesizer (0.32 mmol scale) and Biotage purification gives an oilycompound. Slow addition of HCl (1M in Et₂O) into a solution of the abovecompound in MeOH gives the hydrochloride salt of the title compound(0.085 g) as a white solid in a 63% yield.

Mp: 207° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.12 (d, 2H, J=7.7 Hz), 7.64 (d, 2H,J=8.2 Hz), 7.43 (tr, 2H, J=7.23, 8.1 Hz), 7.31 (d, 2H, J=8.12), 7.19(tr, 2H, J=7.4 Hz), 6.80 (d, 2H, J=8.45 Hz), 4.39 (br s, 2H), 4.17 (brs, 1H), 3.45-3.28 (br m, 12H).

Example 56(±)-1-[4-(4-tert-Butylbenzyl)piperazin-1-yl]-3-(carbazol-9-yl)propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from 4-tert-butylbenzaldehyde using a parallel Quest 210synthesizer (0.32 mmol scale) and Biotage purification gives an oilycompound. Slow addition of HCl (1M in Et₂O) into a solution of the abovecompound in MeOH gives the hydrochloride salt of the title compound(0.085 g) as a white solid in a 57% yield.

Mp: 246° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.12 (d, 2H, J=7.7 Hz), 7.64 (d, 2H,J=8.2 Hz), 7.43 (m, 6H), 7.19 (tr, 2H, J=7.4 Hz), 4.39 (br s, 2H), 4.17(br s, 1H), 3.45-3.28 (br m, 12H), 1.27 (s, 9H).

Example 57(±)-1-(Carbazol-9-yl)-3-[4-(3,4-dichlorobenzyl)piperazin-1yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from 3,4-dichlorobenzaldehyde using a parallel Quest 210synthesizer (0.32 mmol scale) and Biotage purification gives an oilycompound. Slow addition of HCl (1M in Et₂O) into a solution of the abovecompound in MeOH gives the hydrochloride salt of the title compound(0.084 g) as a grey solid in a 56% yield.

Mp: 110° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.12 (d, 2H, J=7.7 Hz), 7.69 (s, 1H),7.67 (m, 3 H), 7.54 (m, 3H), 7.19 (tr, 2H, J=7.4 Hz), 4.39 (br s, 2H),4.17 (br s, 1H), 3.45-3.20 (br m, 12H).

Example 58(±)-1-(Carbazol-9-yl)-3-{4-[4-(methylsulfonyl)benzyl])piperazin-1-yl}propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from 4-(methylsulfonyl)benzaldehyde using a parallel Quest 210synthesizer (0.32 mmol scale) and Biotage purification gave an oilycompound. Slow addition of HCl (1M in Et₂O) into a solution of the abovecompound in MeOH gives the hydrochloride salt of the title compound(0.091 g) as a white solid in a 59% yield.

Mp: 85° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.12 (d, 2H, J=7.7 Hz), 7.90 (d, 2H,J=8.2 Hz), 7.6-7.62 (m, 4H), 7.46-7.40 (tr, 2H, J=8.2 Hz), 7.19 (tr, 2H,J=7.4 Hz), 4.39 (br s, 2H), 4.17 (br s, 1H), 3.45-3.20 (br m, 12H), 3.18(s, 3H).

Example 59(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(thien-2-ylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 5 and thiophene-2-carboxaldehyde using a parallelQuest 210 synthesizer (0.21 mmol scale) and Biotage purification gave anoily compound. Slow addition of HCl (1M in Et₂O) into a solution of theabove compound in MeOH gives the hydrochloride salt of the titlecompound (0.066 g) as an off-white solid in a 57% yield.

Mp: 262° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.54 (d, 2H, J=1.71 Hz), 7.69-7.57 (m,5H), 7.15 (s, 1H), 7.05 (tr, 1H, J=4.69, 3.67 Hz), 4.37 (br s, 2H), 4.25(br s, 1H), 3.52-2.93 (m, 12H).

Example 60(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(thien-3-ylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 5 and thiophene-3-carboxaldehyde using a parallelQuest 210 synthesizer (0.21 mmol scale) and Biotage purification givesan oily compound. Slow addition of HCl (1M in Et₂O) into a solution ofthe above compound in MeOH gives the hydrochloride salt of the titlecompound (0.09 g) as a pale yellow solid in a 76% yield.

Mp: 268-271° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.46 (d, 2H, J=1.82 Hz), 7.74-7.57 (m,6H), 7.28 (d, 1H, J=4.64 Hz), 4.4-4.2 (br s, 2H), 4.1 (dd, 1H, J=5.2,1.7 Hz), 3.52-2.93 (m, 12H).

Example 61(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(pyridin-3-ylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 5 and nicotinaldehyde using a parallel Quest 210synthesizer (0.21 mmol scale) and Biotage purification gives an oilycompound. Slow addition of HCl (1M in Et₂O) into a solution of the abovecompound in MeOH gives the hydrochloride salt of the title compound(0.057 g) as a hygroscopic salmon-colored powder in a 30% yield.

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.8 (m, 2H), 8.47 (2 d, 2H, J=8.3, 1.8Hz), 8.32 (d, 1H, J=7.92 Hz), 8.01 (dd, 1H, J=8.0 Hz), 7.82 (m, 1H),7.69-7.57 (m, 3H), 4.4-4.2 (br s, 2H), 4.1 (dd, 1H, J=5.2, 1.7 Hz),3.52-2.93 (m, 12H).

Example 62(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-methoxybenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 5 and 4-methoxybenzaldehyde using a parallel Quest210 synthesizer (0.21 mmol scale) and Biotage purification gives an oilycompound. Slow addition of HCl (1M in Et₂O) into a solution of the abovecompound in MeOH gives the hydrochloride salt of the title compound(0.053 g) as a white powder in a 43% yield.

Mp: 251° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.45 (d, 2H, J=1.7 Hz), 7.70-7.57 (m,4H), 7.42 (d, 2H, J=7.9 Hz), 6.98 (d, 2H, J=8.48 Hz), 4.4-4.2 (br s,2H), 4.1 (dd, 1H, J=5.2, 1.7 Hz), 3.9 (s, 3H), 3.52-2.93 (m, 12H).

Example 63(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-tert-butylbenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 5 and 4-tert-butyl benzaldehyde using a parallelQuest 210 synthesizer (0.21 mmol scale) and Biotage purification givesan oily compound. Slow addition of HCl (1M in Et₂O) into a solution ofthe above compound in MeOH gives the hydrochloride salt of the titlecompound (0.061 g) as a white powder in a 48% yield.

Mp: 242° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.46 (d, 2H, J=1.7 Hz), 7.68 (d, 2H,J=8.8 Hz), 7.61-7.58 (dd, 2H, J=8.7, 1.85 Hz), 7.47 (br s, 4H), 4.4-4.2(br s, 3H), 3.52-2.93 (m, 12H), 1.27 (s, 9H).

Example 64(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-trifluoromethylbenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 5 and 4-trifluoromethylbenzaldehyde using aparallel Quest 210 synthesizer (0.21 mmol scale) and Biotagepurification gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound (0.058 g) as a pale yellow powder in a 44%yield.

Mp: 243° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.46 (d, 2H, J=1.7 Hz), 7.88-7.50 (m,8H), 4.4-4.2 (br s, 2H), 4.12 (br s, 1H), 3.52-2.93 (m, 12H).

Example 65(±)-1-[4-(1,3-Benzodioxol-5-ylmethyl)piperazin-1-yl]-3-(3,6-dibromo-carbazol-9-yl)propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 5 and 1,3-benzodioxole-5-carboxaldehyde using aparallel Quest 210 synthesizer (0.21 mmol scale) and Biotagepurification gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound (0.051 g) as a pale yellow powder in a 40%yield.

Mp: 272° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.46 (d, 2H, J=1.7 Hz), 7.70-7.57 (m,4H), 7.11 (s, 1H), 6.97 (s, 2H), 6.046 (s, 2H), 4.4-4.2 (br s, 2H), 4.12(dd, 1H, J=5.6, 1.8 Hz), 3.52-2.93 (m, 12H).

Example 66(±)-1-(4-Cyclohexylmethylpiperazin-1-yl)-3-(3-phenylcarbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom Intermediate 7, Intermediate 11 and sodium hydride and heating at60° C. for 16 hours gives after flash chromatography the title compoundas a yellow foam. Slow addition of HCl (1M in Et₂O) into a solution ofthe above compound in DCM gives the hydrochloride salt of the titlecompound as a white solid in a 90% yield.

Mp: 265-270° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.46 (s, 1H), 8.24 (d, 2H, J=7.67 Hz),7.78-7.69 (m, 5H), 7.50-7.45 (m, 3H), 7.32 (tr, 1H, J=7.3 Hz), 7.22 (tr,1H, J=7.4 Hz), 4.4-4.3 (m, 3H), 3.6-3.0 (m, 11H), 2.9 (br s, 1H),1.8-1.5 (m, 6H), 1.2-0.9 (m, 5H).

Analysis for C₃₂H₃₉N₃O. 2HCl. 0.8 H₂O:

Calculated: C, 67.55; H, 7.55; N, 7.38;

Found: C, 67.61; H, 7.82; N, 7.42%.

Example 67(±)-3,6-Dibromo-9-{3-[4-(cyclohexylmethyl)piperazin-1-yl]-2-fluoropropyl}-carbazole

Under an inert atmosphere a solution of Example 45 (0.10 g, 0.18 mmol)in anhydrous DCM (8 mL) is treated with DAST (0.77 mmol). The resultingslurry is allowed to warm up to rt. Tlc monitoring (SiO₂, DCM/MeOH(95/5) shows formation of a few new UV active compounds anddisappearance of the starting material. The reaction mixture is quenchedwith a saturated aqueous solution of potassium carbonate (10 mL).Extraction with DCM (3×20 mL), drying over MgSO₄ and evaporation of thesolvent under reduced pressure gives an oil. Purification on a 2×21 cm²SiO₂ column using Petroleum Ether/EtOAc/MeOH (6/1/0.15) as eluant givesthe title compound (Rf=0.2) as the UV active major product of thereaction. Slow addition of a solution of the above major compound(Rf=0.2) in DCM into a solution of HCl(1M in Et₂O) gives thehydrochloride salt of the title compound as a beige solid in a 52%yield.

Mp: 265-270° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.48 (s, 2H), 7.67 (d, 2H, J=8.9 Hz),7.62 (d, 2H, J=8.9 Hz), 5.35 (d, 1H, J=51.4 Hz), 4.69 (m, 2H), 3.90-3.10(m, 10H), 2.97 (d, 2H, J=5.1 Hz), 1.85-1.55 (m, 6H), 1.30-1.05 (m, 3H),1.05-0.85 (m, 2H).

Example 68(±)-9-{3-[4-(Cyclohexylmethyl)piperazin-1-yl]-2-fluoropropyl}-3-phenyl-carbazole

The same method as employed in the preparation of Example 67 butstarting from Example 66 gives a more polar UV active compound (Rf=0.17using SiO₂, Petroleum Ether/EtOAc/MeOH (6/1/0.15) as eluting solvent).Slow addition of HCl into a solution of the above compound in DCM givesthe hydrochloride salt of the title compound as a beige solid in a 20%yield.

Mp: 255° C. (decomposition).

FI-MS (APCI): m/z observed in a positive mode: 484.2.

¹H NMR (CDCl₃, 300 MHz) of the base form of the title compound 88.28 (d,1H, J=1.5 Hz), 8.12 (d, 1H, J=7.9 Hz), 7.70 (m, 3H), 7.58-7.42 (m, 5H),7.33 (m, 1H), 7.25 (ddd, 1H, J=7.9, 6.4, 1.5 Hz), 5.04 (dm, 1H, J=47.5Hz), 4.67 (ddd, 1H, J=22.2, 15.7, 4.1 Hz), 4.54 (ddd, 1H, J=20.3, 15.7,6.0 Hz), 2.70-2.40 (m, 10H), 2.16 (d, 2H, J=7.2 Hz), 1.81-1.60 (m, 5H),1.49 (m, 1H), 1.31-1.06 (m, 3H), 0.96-0.77 (m, 2H).

Example 69(±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxypropyl]-N-(4-fluorophenyl)piperazine-1-carboxamide

A solution of Example 5 (0.20 g, 0.43 mmol) in DCM (10 mL) is treatedwith DIEA (80 μL, 1 equiv.) and 1-fluoro-4-isocyanatobenzene (0.06 g, 1equiv.). The resulting solution is allowed to stir for 4 hours. Thesolvent is evaporated off. The crude residue is taken up in DCM (5 mL)and is purified via flash chromatography on SiO₂ using DCM/MeOH (95/5)as eluting solvent. A by-product is identified as(±)-2-(3,6-dibromocarbazol-9-yl)-1-({4-[4(4-fluoroanilino)carbonyl]piperazin-1-yl}methyl)ethyl4-fluorophenylcarbamate (Rf=0.45), FI-MS (APCI): m/z observed in apositive mode: 742.4). The major UV active compound (Rf=0.41) isidentified as the title compound. Slow addition of HCl (1M in Et₂O) intoa solution of the above compound (Rf=0.41) in MeOH gives thehydrochloride salt of the title compound (0.125 g) as a white powder ina 48% yield.

Mp: 184° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.40 (br. s, 2H), 7.60 (d, 2H, J=8.3Hz), 7.55 (d, 2H, J=8.3 Hz), 7.34 (m, 2H), 7.01 (m, 2H), 4.33 (br. s,3H), 4.07 (m, 2H), 3.50-2.90 (m, 8H).

Example 70(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(thien-2-ylsulfonyl)piperazin-1-yl]propan-2-ol

A solution of Example 5 (0.20 g, 0.43 mmol) in DMF (7 mL) is treatedwith TEA (60 uL, 1 equiv.) and thiophene-2-sulfonyl chloride (0.078 g, 1equiv.). The resulting solution is allowed to stir for 2 hours. Thesolvent is evaporated. The crude residue is taken up in DCM (5 mL) andis purified via flash chromatography using DCM/MeOH (95/5) as eluant.Slow addition of HCl (1M in Et₂O) into a solution of the purified abovecompound in MeOH gives the hydrochloride salt of the title compound(0.233 g, 0.36 mmol) as a white powder in a 84% yield.

Mp: 252° C. (decomposition).

Analysis for C₂₃H₂₃Br₂N₃O₃S. HCl:

Calculated: C, 42.51; H, 3.72; N, 6.47;

Found: C, 42.26; H, 4.00; N, 6.55%.

Example 71(±)-1-[4-(Benzylsulfonyl)piperazin-1-yl]-3-(3,6-dibromo-carbazol-9yl)propan-2-ol

The same method as employed in the preparation of Example 70 butstarting from phenyl methane sulfonyl chloride gives the hydrochloridesalt of the title compound as a beige-colored solid in a 71% yield.

Mp: 253° C. (decomposition).

Analysis for C₂₆H₂₇Br₂N₃O₃S. HCl:

Calculated: C, 47.47; H, 4.29; N, 6.39;

Found: C, 47.18; H, 4.31; N, 6.32%.

Example 72(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(3,4-dichlorobenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 5 and 3,4-dichlorobenzaldehyde gives thehydrochloride salt of the title compound as a white powder in a 77%yield.

Mp: 287° C. (decomposition).

Analysis for C₂₆H₂₅Br₂Cl₂N₃O. 2HCl. 0.2H₂O:

Calculated: C, 44.44; H, 3.93; N, 5.98;

Found: C, 44.13; H, 4.08; N, 5.92%.

Example 73(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol

A solution of Intermediate 15 (1.65 g, 4.33 mmol) and4-(2-piperazin-1-ylethyl)morpholine (2.55 g, 12.79 mmol) in anhydrousTHF/absolute EtOH (22.5 mL/22.5 mL) is allowed to stir at 60° C. for 18hours. The solvents are evaporated. The residue is taken up inEtOAc/MeOH/aqueous ammonia (25%) (8/1.5/0.5) and purified on a 5×27 cm²column of SiO₂ using EtOAc/MeOH/aqueous ammonia (25%) (8/1.5/0.5) aseluant to give the title compound (2.45 g, 4.22 mmol) as a pale yellowfoam in a 97% yield. Slow addition of HCl (21.5 mL, 1M in Et₂O) into asolution of the above compound in MeOH/DCM (100 mL/50 mL) gives thehydrochloride of the title compound as a white powder.

Mp: 270° C. (decomposition).

¹H NMR (DMSO-d₆+MeOD-d₄, 300 MHz) δ 8.47 (d, 2H, J=1.9 Hz), 7.72 (d, 2H,J=8.7 Hz), 7.60 (dd, 2H, J=8.7, 1.9 Hz), 4.42 (br s, 3H), 3.88 (br s,4H), 3.75-2.80 (m, 18H).

Analysis for C₂₅H₃₂Br₂N₄O₂. 3HCl:

Calculated: C, 43.53; H, 5.11; N, 8.12;

Found: C, 43.36; H, 5.29; N, 8.06%.

Example 74(±)-1-[4-(4-Fluorobenzyl)piperazin-1-yl]-3-(3-thien-2-yl-carbazol-9-yl)-propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom Intermediate 16, Intermediate 6 and sodium hydride and heating at60° C. for 16 hours gives after flash chromatography the title compoundas a yellow oil. Slow addition of HCl (1M in Et₂O) into a solution ofthe above compound in DCM gives the hydrochloride salt of the titlecompound as a gummy green solid in a 62% yield.

FI-MS (APCI): m/z observed in a positive mode: 500.4.

Example 75(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-{4-[3,4-diethoxyphenyl)sulfonyl]piperazin-1-yl}propan-2-ol

The same method as employed in the preparation of Example 70 butstarting from 3,4-dimethoxybenzenesulfonyl chloride gives thehydrochloride salt of the title compound as a beige solid (0.117 g) in a84% yield.

Mp: 267° C. (decomposition).

Analysis for C₂₇H₂₉Br₂N₃O₅S. HCl:

Calculated: C, 46.07; H, 4.30; N, 5.97;

Found: C, 45.95; H, 4.40; N, 5.88%.

Example 76(±)-1-[4-(Cyclohexylmethyl)piperazin-1-yl]-3-(3,6-dichloro-carbazol-9-yl)propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dichloro-9H-carbazole, Intermediate 11 and sodium hydride andheating at 60° C. for 16 hours gives after flash chromatography thetitle compound (UV active, Rf=0.25 using DCM/MeOH (100/5) as eluant) asa white foam. Slow addition of HCl (1M in Et₂O) into a solution of theabove compound in DCM gives the hydrochloride salt of the title compoundas a white solid in a 71% yield.

Mp: 302° C. (decomposition).

Analysis for C₂₆H₃₃Cl₂N₃O. 2HCl. 0.3 H₂O:

Calculated: C, 56.49; H, 6.49; N, 7.60;

Found: C, 56.48; H, 6.52; N, 7.60%.

Example 77(±)-4-[3-(3,6-Dichlorocarbazol-9-yl)-2-hydroxypropyl]-piperazine-1-carboxylictert-butyl ester

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dichloro-9H-carbazole and(±)-4-oxiranylmethyl-piperazine-1-carboxylic acid tert-butyl ester givesafter flash chromatography the title compound as a white foam in a 74%yield.

¹H NMR (CDCl₃, 300 MHz) δ 7.90 (d, 2H, J=1.32 Hz), 7.39-7.34 (m, 4H),4.32-4.07 (m, 3H), 3.35 (br s, 4H), 2.52-2.22 (m, 6H), 1.41 (s, 9H).

Example 78(±)-1-(3,6-Dichlorocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom Example 77 gives the hydrochloride salt of title compound as awhite solid in a 71% yield.

Mp: 305° C. (decomposition).

Analysis for C₁₉H₂₁Cl₂Br₂N₃O. 2HCl. 0.3H₂O:

Calculated: C, 50.18; H, 5.19; N, 9.24;

Found: C, 50.06; H, 5.15; N, 9.12%.

Example 79(±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-oxopropyl]-piperazine-1-carboxylictert-butyl ester

At −78° C. a solution of oxalyl chloride (200 μL, 2.34 mmol) inanhydrous DCM (20 mL) is treated with anhydrous DMSO (310 μL, 4.37mmol). After 15 min of stirring at −78° C., a solution of Example 4 (1.0g, 1.76 mmol) in anhydrous DCM (5 mL) is added dropwise. The resultingmixture is allowed to stir for 40 min at −78° C. and TEA (1250 μL) isadded neat. After 10 min the reaction mixture is allowed to warm up to−30° C. After 1 hour of stirring at −30° C. the reaction mixture isquenched with water (50 mL). Extraction with DCM (3×100 mL), drying overMgSO₄ and evaporation under reduced pressure gives a yellow oil. Flashchromatography on a 4×21 cm² SiO₂ column using Et₂O/MeOH (100/1) mixturethen (100/3) as eluant gives the title compound as a pale yellow foam(0.804 g, 1.42 mmol) in a 81% yield.

¹H NMR (CDCl₃, 300 MHz) δ 8.12 (d, 2H, J=1.9 Hz), 7.52 (dd, 2H, J=8.6,1.9), 7.12 (d, 2H, J=8.6), 5.00 (s, 2H), 3.39 (m, 4H), 3.05 (s, 2H),2.32 (m, 4H), 1.42 (s, 9H).

Example 80(±)-3,6-Dibromo-9-(2,2-difluoro-3-piperazin-1-ylpropyl)-carbazole

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3,6-dibromocarbazol-9-yl)-2,2-difluoropropyl]piperazine-1-carboxylictert-butylester gives the hydrochloride salt of title compound as abeige solid in a 71% yield.

Mp: 305° C. (decomposition).

FI-MS (APCI): m/z observed in a positive mode: 488.0

Preparation of(±)-4-[3-(3,6-dibromocarbazol-9-yl)-2,2-difluoropropyl]-piperazine-1-carboxylictert-butyl ester

DAST (100 uL, 0.76 mmol) is added into a solution of from(±)-4-[3-(3,6-dibromocarbazol-9-yl)-2-oxopropyl]-piperazine-1-carboxylictert-butylester (0.100 g, 0.18 mmol) in anhydrous DCM (4 mL) at rt. Theresulting mixture is allowed to stir for 21 hours and quenched with asaturated aqueous solution of sodium hydrogenocarbonate (20 mL).Extraction with DCM (3×40 mL), drying over MgSO₄ and evaporation underreduced pressure gives a brown oil. Flash chromatography on a 2×21 cm²SiO₂ column using DCM/MeOH (100/1.5) as eluant gives the title compoundas a yellow oil.

¹H NMR (CDCl₃, 300 MHz) δ 8.09 (d, 2H, J=1.9 Hz), 7.53 (dd, 2H, J=8.7,1.9 Hz), 7.42 (d, 2H, J=8.7 Hz), 4.67 (t, 2H, J=13.0 Hz), 3.47 (m, 4H),2.64 (t, 2H, J=13.0 Hz), 2.49 (br. s, 4H), 1.46 (s, 9H).

Example 81(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-phenylethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and phenyl acetaldehyde using a parallel Radleysynthesizer (0.21 mmol scale) and parallel flash purification (ISCOdevice) gives an oily compound. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in MeOH gives the hydrochloride salt ofthe title compound as a beige solid in a 62% yield.

Mp: 264° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.39 (d, J=1.88 Hz, 1H), 8.20 (d,J=7.54 Hz, 1H), 7.71 (tr, J=9.04, 9.42 Hz, 2H), 7.57 (dd, J=8.67, 1.9Hz, 1H), 7.45 (tr, J=7.91, 7.54 Hz, 1H), 7.35-7.20 (m, 6H), 4.42 (br s,3H), 4.03-3.07 (m, 14H).

Example 82(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2,3-dihydro-1,4-benzodioxin-6-ylmethyl)piperazin-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and2,3-dihydro-1,4-dihydro-1,4-benzodioxine-6-carboxaldehyde using aparallel Radley, synthesizer (0.21 mmol scale) and parallel flashpurification (ISCO device) gives an oily compound. Slow addition of HCl(1M in Et₂O) into a solution of the above compound in MeOH gives thehydrochloride salt of the title compound as a beige solid in a 49%yield.

Mp: 230° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.38 (br s, 1H), 8.19 (d, 1H, J=7.54Hz), 7.71 (tr, 2H, J=9.04, 9.42 Hz), 7.57 (dd, 1H, J=8.67, 1.9 Hz), 7.45(tr, 1H, J=7.91, 7.54 Hz), 7.24 (tr, 1H, J=7.53 Hz), 7.14 (br s, 1H),7.02 (br s, 1H), 6.90 (d, 1H, J=7.91 Hz), 4.39 (br s, 3H), 4.23 (s, 2H),4.015 (s, 2H), 4.49-3.3.27 (m, 12H).

Example 83(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(4-fluorobenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 4-fluorobenzaldehyde using a parallel Radleysynthesizer (0.21 mmol scale) and parallel flash purification (ISCOdevice) gives an oily compound. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in MeOH gives the hydrochloride salt ofthe title compound as a white solid in a 67% yield.

Mp: 267° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.38 (d, 1H, J=1.88 Hz), 8.19 (d, 1H,J=7.54 Hz), 7.73-7.67 (m, 3H), 7.57 (dd, 1H, J=8.67, 1.9 Hz), 7.45 (tr,1H, J=7.91, 7.54 Hz), 7.32-7.20 (m, 4H), 7.14 (br s, 1H), 4.40 (br s,3H), 3.64-3.28 (m, 12H).

Example 84(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(3,4-dichlorobenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 3,4-dichlorobenzaldehyde using a parallelRadley synthesizer (0.21 mmol scale) and parallel flash purification(ISCO device) gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound as a white solid in a 52% yield.

Mp: 183° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.38 (d, 1H, J=1.88 Hz), 8.19 (d, 1H,J=7.54 Hz), 7.88 (br s, 1H), 7.72-7.67 (m, 3H), 7.57 (dd, 2H, J=8.67,1.9 Hz), 7.45 (tr, 1H, J=7.91, 7.54 Hz), 7.21 (tr, 1H, J=7.53, 7.0 Hz),4.40 (br s, 3H), 3.64-3.28 (m, 12H).

Example 85(±)-1-(3-Bromo-carbazol-9-yl)-3-{4-[4-(difluoromethoxybenzyl]piperazin-1-yl}propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 4-difluoromethoxy benzaldehyde using aparallel Radley synthesizer (0.21 mmol scale) and parallel flashpurification (ISCO device) gives an oily compound. Slow addition of HCl(1M in Et₂O) into a solution of the above compound in MeOH gives thehydrochloride salt of the title compound as a white solid in a 37%yield.

Mp: 184° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.38 (d, 1H, J=1.88 Hz), 8.19 (d, 1H,J=7.54 Hz), 7.77-7.66 (m, 4H), 7.57 (dd, 1H, J=8.67, 1.9 Hz), 7.45 (tr,1H, J=7.91, 7.54 Hz), 7.21 (m, 4H), 4.40 (br s, 3H), 3.64-3.28 (m, 12H).

Example 86(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(cyclohexylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and cyclohexanecarboxaldehyde using a parallelRadley synthesizer (0.21 mmol scale) and parallel flash purification(ISCO) gives an oily compound. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in MeOH gives the hydrochloride salt ofthe title compound as a white solid in a 48% yield.

Mp: 271° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.39 (d, 1H, J=1.88 Hz), 8.20 (d, 1H,J=7.54 Hz), 7.74-7.68 (tr, 2H, J=9.42, 8.66 Hz), 7.57 (dd, 1H, J=8.17,1.88 Hz), 7.48 (tr, 1H, J=7.54 Hz), 7.22 (tr, 1H, J=7.54, 7.16 Hz), 5.35(d, 1H, J=51.4 Hz), 4.69 (m, 2H), 3.90-3.10 (m, 10H), 2.97 (d, 2H, J=5.1Hz), 1.85-1.55 (m, 6H), 1.30-1.05 (m, 3H), 1.05-0.85 (m, 2H).

Example 87(±)-1-[4-(1,3-Benzodioxol-5ylmethyl)piperazin-1-yl]-3-(3-bromo-carbazol-9-yl)propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 1,3-benzodioxole-5-carboxaldehyde using aparallel Radley synthesizer (0.21 mmol scale) and parallel flashpurification (ISCO device) gives an oily compound. Slow addition of HCl(1M in Et₂O) into a solution of the above compound in MeOH gives thehydrochloride salt of the title compound as a white solid in a 49%yield.

Mp: 199° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.39 (d, 1H, J=1.88 Hz), 8.20 (d, 1H,J=7.54 Hz), 7.68 (m, 2H), 7.57-7.44 (m, 2H), 7.24 (m, 2H), 6.97 (m, 2H),6.046 (s, 2H), 4.4-4.2 (br s, 2H), 4.12 (dd, 1H, J=5.6, 1.8 Hz),3.52-2.93 (m, 12H).

Example 88(±)-1-(3-Bromocarbazol-9-yl)-3-[4-(4-methoxybenzy)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 4-methoxybenzaldehyde using a parallelRadley synthesizer (0.21 mmol scale) and parallel flash purification(ISCO device) gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound as a white solid in a 68% yield.

Mp: 212° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.38 (d, 1H, J=1.88 Hz), 8.20 (d, 1H,J=7.54 Hz), 7.68 (m, 2H), 7.57-7.44 (m, 4H), 7.24 (tr, 1H, J=7.53, 7.16Hz), 6.9 (d, 2H, J=8.28 Hz), 4.4-4.2 (br s, 2H), 4.12 (dd, 1H, J=5.6,1.8 Hz), 3.52-2.93 (m, 15H).

Example 89(±)-1-(3-Bromocarbazol-9-yl)-3-[4-(4-trifluoromethylbenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 4-trifluoromethyl benzaldehyde using aparallel Radley synthesizer (0.21 mmol scale). Parallel flashpurification (ISCO device) gives an oily compound. Slow addition of HCl(1M in Et₂O) into a solution of the above compound in MeOH gives thehydrochloride salt of the title compound as a white solid in a 61%yield.

Mp: 177° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.38 (br s, 1H), 8.20 (d, 1H, J=7.54Hz), 7.80-7.65 (m, 6H), 7.57-7.44 (d, 1H, J=8.67 Hz), 7.50-7.44 (tr, 1H,J=7.54 Hz), 7.20 (tr, 1H, J=7.54, 7.16 Hz), 4.4-4.2 (br s, 3H),3.52-2.93 (m, 12H).

Example 90(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(3,5-dichlorobenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 3,5-dichlorobenzaldehyde using a parallelRadley synthesizer (0.21 mmol scale) and parallel flash purification(ISCO device) gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound as a white solid in a 55% yield.

Mp: 259° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.38 (d, 1H, J=1.88 Hz), 8.19 (d, 1H,J=7.54 Hz), 7.72-7.65 (m, 4H), 7.57 (dd, 2H, J=8.67, 1.9 Hz), 7.45 (tr,1H, J=7.91, 7.54 Hz), 7.21 (tr, 1H, J=7.53, 7.0 Hz), 4.40 (br s, 3H),3.64-3.28 (m, 12H).

Example 91(±)-1-(3-Bromocarbazol-9-yl)-3-[4-(4-tert-butylbenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 4-tert-butylbenzaldehyde using a parallelRadley synthesizer (0.21 mmol scale) and parallel flash purification(ISCO device) gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound as a white solid in a 49% yield.

Mp: 250° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.38 (br s, 1H), 8.19 (d, 1H, J=7.54Hz), 7.72-7.65 (tr, 2H, J=8.66 Hz), 4H), 7.57-7.44 (m, 6H), 7.21 (tr,1H, J=7.53, 7.0 Hz), 4.40 (br s, 3H), 3.64-3.28 (m, 12H), 1.27 (s, 9H).

Example 92(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-furylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 2-furaldehyde using a parallel Radleysynthesizer (0.21 mmol scale) and parallel flash purification (ISCOdevice) gives an oily compound. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in MeOH gives the hydrochloride salt ofthe title compound as a yellow solid in a 34% yield.

Mp: 248° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.38 (br s, 1H), 8.19 (d, 1H, J=7.92Hz), 7.79 (br s, 1H), 7.66 (m, 2H), 7.55 (d, 1H, J=8.1 Hz), 7.47 (tr,1H, J=7.54, 7.16 Hz), 7.22 (tr, 1H, J=7.53, 7.16 Hz), 6.68 (br s, 1H),6.54 (br s, 1H), 4.40 (br s, 3H), 3.64-3.28 (m, 12H).

Example 93(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-furylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 5-methyl-2-furaldehyde using a parallelRadley synthesizer (0.21 mmol scale) and parallel flash purification(ISCO device) gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound as a white solid in a 67% yield.

Mp: 240° C. (decomposition).

¹H NMR (DMSO-d₆, 300 MHz) δ 8.38 (br s, 1H), 8.19 (d, 1H, J=7.92 Hz),7.66 (m, 2H), 7.55 (d, 1H, J=8.1 Hz), 7.47 (tr, 1H, J=7.54, 7.16 Hz),7.22 (tr, 1H, J=7.53, 7.16 Hz), 6.68 (br s, 1H), 6.54 (br s, 1H), 4.40(br s, 3H), 3.64-3.28 (m, 12H), 2.26 (s, 3H).

Example 94(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-pyridin-ylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and pyridine-2-carboxaldehyde using a parallelRadley synthesizer (0.21 mmol scale) and parallel flash purification(ISCO device) gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound as a pale beige solid in a 68% yield.

Mp: 240° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.71 (d, 1H, J=4.15 Hz), 8.38 (s, 1H),8.22 (d, 1H, J=7.54 Hz), 8.69 (tr, 1H, J=7.54, 7.16 Hz), 7.74 (m, 3H),7.63-7.54 (m, 2H), 7.47 (tr, 1H, J=7.54 Hz), 7.22 (tr, 1H, J=7.53, 7.16Hz), 4.40 (br s, 3H), 4.12 (br s, 2H), 3.64-3.28 (m, 10H).

Example 95(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(3-pyridin-ylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and nicotinaldehyde using a parallel Radleysynthesizer (0.21 mmol scale) and parallel flash purification (ISCOdevice) gives an oily compound. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in MeOH gives the hydrochloride salt ofthe title compound as a white solid in a 73% yield.

Mp: 183° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.95 (s, 1H), 8.84 (d, 1H, J=4.9 Hz),8.50 (d, 1H, J=7.91 Hz), 8.38 (d, 1H, J=1.13 Hz), 8.19 (d, 1H J=7.92Hz), 7.90 (tr, 1H, J=6.79, 6.4 Hz), 7.70 (m, 2H), 7.57-7.44 (m, 2H),7.20 (tr, 1H, J=7.53, 7.16 Hz), 4.41-4.29 (br s, 7H), 3.64-3.28 (m, 8H).

Example 96(±)-1-(3-Bromocarbazol-9-yl)-3-[4-(4-pyridin-ylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and isonicotinaldehyde using a parallel Radleysynthesizer (0.21 mmol scale) and parallel flash purification (ISCOdevice) gives an oily compound. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in MeOH gives the hydrochloride salt ofthe title compound as a pale beige solid in a 87% yield.

Mp: 170° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.87 (m, 2H), 8.39 (d, 1H, J=1.89 Hz),8.21 (d, 1H, J=7.91 Hz), 7.99 (m, 2H), 7.39-7.53 (m, 3H), 7.47 (tr, 1H,J=7.53 Hz), 7.20 (tr, 1H, J=7.53, 7.16 Hz), 4.41-4.29 (br s, 7H),3.64-3.28 (m, 8H).

Example 97(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(quinolin-2-ylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and quinoline-2-carboxaldehyde using a parallelRadley synthesizer (0.21 mmol scale) and parallel flash purification(ISCO device) gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound as an orange solid in a 67% yield.

Mp: 167° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.70-7.6 (m, 10H), 7.6-7.4 (m, 2H),7.20 (tr, 1H, J=7.53, 7.16 Hz), 4.41-4.29 (br s, 5H), 3.64-3.28 (m,10H).

Example 98(±)-1-(3-Bromo-carbazol-9-yl)-3-[4-(2-furyl-4-bromomethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 4-bromo-2-furaldehyde using a parallelRadley synthesizer (0.21 mmol scale) and parallel flash purification(ISCO device) gives an oily compound. Slow addition of HCl (1M in Et₂O)into a solution of the above compound in MeOH gives the hydrochloridesalt of the title compound as a beige solid in a 68% yield.

Mp: 245° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.38 (d, 1H, J=1.88 Hz), 8.19 (d, 1H,J=7.53 Hz), 8.03 (s, 1H), 7.69 (m, 2H), 7.55 (dd, 1H, J=8.8, 2.0 Hz),7.47 (tr, 1H, J=7.15 Hz), 7.20 (tr, 1H, J=7.54, 7.16 Hz), 6.83 (s, 1H),4.39 (br s, 2H), 4.09 (m, 6H), 3.62-3.10 (br m, 7H).

Example 99(±)-1-(3-Bromocarbazol-9-yl)-3-[4-(1-naphtylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 1-naphtaldehyde using a parallel Radleysynthesizer (0.21 mmol scale) and parallel flash purification (ISCOdevice) gives an oily compound. Slow addition of HCl(1M in Et₂O) into asolution of the above compound in MeOH gives the hydrochloride salt ofthe title compound as a white solid in a 25% yield.

Mp: 185° C. (decomposition).

¹H NMR (DMSO-d₆, 300 MHz) δ 8.38 (m, 2H), 8.19 (d, 1H, J=7.54 Hz), 8.03(br s, 2H), 7.8 7.44 (m, 8H), 7.20 (tr, 1H, J=7.54, 7.16 Hz), 4.39-3.0(br m, 15H).

Example 100(±)-1-{4-[(6-Bromo-1,3-benzodioxol-5-yl)methyl]piperazin-1-yl}-3-(3-bromo-carbazol-9-yl)propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 6-bromo-1,3-dioxole-5-carboxaldehyde using aparallel Radley synthesizer (0.21 mmol scale) and parallel flashpurification (ISCO device) gives an oily compound. Slow addition of HCl(1M in Et₂O) into a solution of the above compound in MeOH gives thehydrochloride salt of the title compound as a white solid in a 58%yield.

Mp: 265° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD 300 MHz) δ 8.38 (d, 1H, J=1.89 Hz), 8.20 (d, 1H,J=7.91 Hz), 7.70 (br tr, 2H), 7.56 (dd, 1H, J=8.8, 1.7 Hz), 7.49 (tr,1H, J=7.54 Hz), 7.38 (br s, 1H), 7.30 (s, 1H), 7.20 (tr, 1H, J=7.54 Hz),6.11 (s, 2H), 4.40 (br s, 3H), 4.09 (br s, 5H), 3.62-3.14 (m, 7H)

Example 101(±)-1-{4-[(6-Chloro-1,3-benzodioxol-5-yl)methyl]piperazin-1-yl}-3-(3-bromo-carbazol-9-yl)propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and 6-chloro-1,3-dioxole-5-carboxaldehyde usinga parallel Radley synthesizer (0.21 mmol scale) and parallel flashpurification (ISCO device) gives an oily compound. Slow addition of HCl(1M in Et₂O) into a solution of the above compound in MeOH gives thehydrochloride salt of the title compound as a white solid in a 82%yield.

Mp: 273° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 8.38 (d, 1H, J=1.89 Hz), 8.20 (d, 1H,J=7.91 Hz), 7.70 (br tr, 2H), 7.56 (dd, 1H, J=8.8, 1.7 Hz), 7.49 (tr,1H, J=7.54 Hz), 7.38 (br s, 1H), 7.30 (s, 1H), 7.20 (br s, 2H), 6.11 (s,2H), 4.40 (br s, 3H), 4.09 (br s, 5H), 3.62-3.14 (m, 7H)

Example 102(±)-1-(3-Chlorocarbazol-9-yl)-3-[4-(4-fluorobenzyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 73 butstarting from Intermediate 17 and 1-(4-fluorobenzyl)piperazine gives thehydrochloride salt of the title compound as a white solid in a 27%yield.

Mp: 258.3° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.26 (d, 1H, J=1.88 Hz), 8.19 (d, 1H,J=7.54 Hz), 7.73-7.61 (m, 4H), 7.48-7.43 (m, 2H), 7.32-7.22 (m, 3H),4.40 (br s, 2H), 4.13 (br s, 1H), 3.46-3.30 (m, 12H).

Example 103(±)-1-(3-Chlorocarbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 73 butstarting from Intermediate 17 gives the hydrochloride salt of the titlecompound as a white solid in a 57% yield.

Mp: 250.7° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.26 (d, 1H, J=1.84 Hz), 8.20 (d, 1H,J=7.53 Hz), 7.72 (m, 2H), 7.50 (m, 2H), 7.23 (tr, 2H, J=7.53, 7.14 Hz),4.42 (br s, 3H), 3.88 (br s, 4H), 3.75-2.80 (m, 18H).

Example 104(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(3-piperidin-1-yl-propyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 1 but startingfrom 3,6-dibromo-9H-carbazole, Intermediate 18 and sodium hydride andheating at 60° C. for 16 hours gives after flash chromatography thetitle compound as a white foam. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in DCM gives the hydrochloride salt ofthe title compound as a beige solid in a 53% yield.

Mp: 267° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.45 (d, 2H, J=1.9 Hz), 7.68 (d, 2H,J=8.8 Hz), 7.60 (dd, 2H, J=8.7, 1.9 Hz), 4.4-4.3 (m, 3H), 3.5-2.6 (m,18H), 1.9 (m, 2H), 1.4 (m, 4H), 1.3 (m, 2H).

Example 105(±)-1-(3-Chlorocarbazol-9-yl)-3-(4-cyclohexylpiperazin-1-yl)propan-2-ol

A solution of NaH (0.026 g, 0.60 mmol, 55% in mineral oil) is added intoa solution of 3-chloro-9H-carbazole (0.100 g, 0.50 mmol) in anhydrousTHF (5 mL). After 30 min of stirring at rt a solution of(±)-3-nitro-benzenesulfonic acid oxiranylmethyl ester in anhydrous THF(0.5 mL) is added. After 90 min of stirring at rt a solution of1-cyclohexyl piperazine (0.25 g, 1.50 mmol) in anhydrous THF (1 mL) isthen added. The resulting mixture is allowed to stir at 60° C. for 15hours. The reaction mixture is quenched with brine (15 mL) and extractedwith Et₂O (50 mL+2×25 mL). After drying over MgSO₄, evaporation underreduced pressure, the crude compound is purified via flashchromatography on a 3×24 cm² SiO₂ column using EtOAc/MeOH (8/1) and(4/1) as eluant. A solution of the purified above compound (0.088 g,0.21 mmol, 41%) is treated with HCl (1M in Et₂O) to give the titlecompound hydrochloride (0.085 g, 0.17 mmol) as a white powder in a 34%yield.

Mp: 316° C. (decomposition).

¹H NMR (DMSO-d₆+MeOD-d₄ (35:1), 300 MHz) δ 8.25 (d, 1H, J=2.0 Hz), 8.19(d, 1H, J=7.9 Hz), 7.74 (d, 1H, J=8.7 Hz), 7.72 (d, 1H, J=7.9 Hz), 7.48(dd, 1H, J=7.9, 7.5 Hz), 7.45 (dd, 1H, J=8.7, 2.0 Hz), 7.22 (dd, 1H,J=7.9, 7.5 Hz), 4.42 (br s, 3H), 4.20-3.10 (m, 11H), 2.06 (br d, 2H,J=9.4 Hz), 1.81 (br d, 2H, J=12.1 Hz), 1.59 (br d, 1H, J=11.7 Hz),1.50-1.00 (m, 5H).

Example 106(±)-1-(3-Bromocarbazol-9-yl)-3-[4-(quinolin-4-ylmethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 47 butstarting from Example 9 and quinoline-4-carboxaldehyde using a parallelRadley synthesizer (0.21 mmol scale). Parallel flash purification (ISCOdevice) gives an oily compound. Slow addition of HCl (1M in Et₂O) into asolution of the above compound in MeOH gives the hydrochloride salt ofthe title compound as a pale yellow solid in a 73% yield.

Mp: 192° C. (decomposition).

¹H NMR (DMSO-d₆+CD₃OD, 300 MHz) δ 9.221 (d, 1H, J=5.28 Hz), 8.53 (d, 1H,J=8.29 Hz), 8.37 (br s, 2H), 8.19 (d, 1H, J=7.53 Hz), 8.07 (m, 2H), 7.89(tr, 1H, J=7.54 Hz), 7.71 (tr, 2H, J=9.05, 8.66 Hz), 7.50 (d, 1H, J=8.67Hz), 7.46 (tr, 1H, J=7.54 Hz), 7.24 (tr, 1H, J=7.54, 7.16 Hz), 4.99 (brs, 5H), 3.48-2.93 (m, 10H).

Analysis for C₂₉H₂₉BrN₄O. 2HCl. 2.1H₂O:

Calculated: C, 54.41; H, 5.54; N, 8.75;

Found: C, 54.45; H, 5.41; N, 8.64%:

Example 107(±)-4-[3-(3-Chlorocarbazol-9-yl)-2-hydroxypropyl]-3,5-dimethylpiperazine-1-carboxylictert-butyl ester

The same method as employed in the preparation of Example 73 butstarting from Intermediate 17 and 3,5-dimethylpiperazine-1-carboxylictert-butyl ester gives after flash chromatography the title compound asa white solid in a 85% yield.

¹H NMR (CDCl₃, 300 MHz) δ 8.13-7.92 (m, 2H), 7.58-7.35 (m, 3H),7.34-7.16 (m, 2H), 4.43-4.21 (m, 2H), 4.17-3.94 (1H, m), 3.9-3.6 (m,1H), 3.51 (bs, 1H), 2.8-2.36 (m, 4H), 1.72-1.49 (m, 3H), 1.42 (s, 9H),1.05-0.8 (m, 6H).

Example 108(±)-1-[4-(cyclohexylmethyl)piperazin-1-yl]-3-(3,6-dichlorocarbazol-9-yl)acetone

At −78° C. a solution of oxalyl chloride (28 uL, 0.328 mmol) inanhydrous DCM (2 mL) is treated with anhydrous DMSO (43 uL, 0.613 mmol).After 15 min of stirring at −78° C., a solution of Example 76 (0.078 g,0.164 mmol) in anhydrous DCM (0.5 mL) is added dropwise. The resultingmixture is allowed to stir for 40 min at −78° C. and TEA (125 μL) isadded neat. After 10 min the reaction mixture is allowed to warm up to−30° C. After 1 hour of stirring at −30° C. the reaction mixture isquenched with water (15 mL). Extraction with DCM (3×25 mL), drying overMgSO₄ and evaporation under reduced pressure gives a yellow oil. Flashchromatography on a 3×25 cm² SiO₂ column using DCM/MeOH (100/5) mixturethen (100/3) as eluting solvent gives the title compound as a paleyellow foam (0.045 g) in a 58% yield. Slow addition of HCl (1M in Et2O)is added to a solution of the above compound in DCM to give thehydrochloride salt of the title compound as a white powder.

Mp: 220° C. (decomposition).

¹H NMR (DMSO-d₆+MeOD-d₄ (35:1), 300 MHz) δ 8.34 (br. s, 2H), 7.60 (d,2H, J=8.7 Hz), 7.48 (br. d, 2H, J=8.7 Hz), 5.55 (br. s, 2H), 4.31 (br.s, 2H), 3.60 (br. s, 2H), 3.32 (br. s, 6H), 2.96 (br. s, 2H), 1.90-1.53(m, 6H), 1.31-1.07 (m, 3H), 1.03-0.84 (m, 2H).

Example 109(±)-1-[4-(cyclohexylmethyl)piperazin-1-yl]-3-(3,6-dibromocarbazol-9-yl)acetone

The same method as employed in the preparation of Example 108 butstarting from Example 45 gives the hydrochloride salt of the titlecompound as a white solid in a 64% yield.

Mp: 261° C. (decomposition).

Analysis for C₂₆H₃₁Br₂N₃O. 2HCl. 0.7H₂O:

Calculated: C, 48.27; H, 5.36; N, 6.50;

Found: C, 48.29; H, 5.50; N, 6.47%:

Example 110(±)-1-(3,6-Dichlorocarbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 73 butstarting from (±)-3,6-dichloro-9-(oxiran-2-ylmethyl)-9H-carbazole givesthe hydrochloride salt of the title compound as a white solid in a 30%yield.

Mp: 253° C. (decomposition).

Analysis for C₂₂H₃₂Cl₂N₄O₂. 3HCl. 0.3H₂O:

Calculated: C, 49.53; H, 5.92; N, 9.24;

Found: C, 49.56; H, 5.91; N, 9.19%:

Example 111(±)-1-[4-(Cyclohexylmethyl)piperazin-1-yl]-3-(3-phenylcarbazol-9-yl)acetone

The same method as employed in the preparation of Example 108 butstarting from Example 68 gives the hydrochloride salt of the titlecompound as a beige solid in a 76% yield.

Mp: 231° C. (decomposition).

Analysis for C₃₂H₃₇N₃O. 2HCl. 0.4H₂O:

Calculated: C, 68.66; H, 7.17; N, 7.51;

Found: C, 68.69; H, 7.01; N, 7.47%:

Example 112(±)-1-(3-Bromocarbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 105 butstarting from 3-bromo-9H-carbazole and4-(2-piperazin-1-ylethyl)morpholine gives the hydrochloride salt of thetitle compound as a white powder in a 38% yield.

Mp: 248° C. (decomposition).

¹H NMR (DMSO-d₆+MeOD-d₄ (35:1), 300 MHz) δ 8.39 (br s, 1H), 8.20 (d, 1H,J=7.5 Hz), 7.73 (d, 1H, J=8.7 Hz), 7.70 (d, 1H, J=7.9 Hz), 7.56 (br d,1H, J=8.7 Hz), 7.48 (dd, 1H, J=7.9, 7.5 Hz), 7.22 (dd, 1H, J=7.5, 7.5Hz), 4.42 (m, 3H), 3.88 (br s, 4H), 3.75-2.70 (m, 18H).

Example 113(±)-1-(3-Chloro-carbazol-9-yl-)-3-(3,5-dimethylpiperazine-1-yl)propan-2-ol

The same method as employed in the preparation of Example 73 butstarting from Intermediate 17 and 2,6-dimethylpiperazine gives thehydrochloride salt of the title compound as a beige solid in a 99%yield.

Mp: 65° C. (decomposition).

¹H NMR (CDCl₃, 300 MHz) δ 8.11-7.96 (m, 2H), 7.55-7.35 (m, 4H),7.31-7.18 (m, 1H), 5.50 (br s, 2H), 4.34 (dd, 2H, J=4.90, 2.26 Hz),4.25-4.14 (m, 1H), 2.97-2.69 (m, 3H), 2.67-2.53 (m, 1H), 2.47-2.33 (m,2H), 1.87 (t, 1H, J=10.36 Hz), 1.55 (t, 1H, J=10.17 Hz), 0.99 (dd, 6H,J=5.84, 0.94 Hz)

Example 114(±)-1-(3-Chlorocarbazol-9-yl-)-3-(2,6-dimethylpiperazin-1-yl)propan-2-ol

The same method as employed in the preparation of Example 3 but startingfrom Example 107 gives the title compound in a 84% yield as a beigepowder.

Mp: 204° C. (decomposition).

¹H NMR (CDCl₃, 300 MHz) δ 8.10-7.99 (m, 2H), 7.55-7.35 (m, 4H),7.30-7.20 (m, 1H), 5.37 (br s, 2H), 4.31 (dd, 2H, J=5.46, 2.07 Hz),4.15-4.00 (m, 1H), 2.91-2.61 (m, 3H), 2.6-2.32 (m, 5H), 0.91 (dd, 6H,J=16.6, 5.65 Hz).

Example 115(±)-1-(3,6-Dibromo-carbazol-9-yl-)-3-piperazin-1-ylpropan-2-amine

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3,6-dibromocarbazol-9-yl)-2-aminopropyl]-piperazine-1-carboxylictert-butyl ester gives the title compound in a 99% yield as a colorlessoil. Slow addition of HCl (1M) into a solution of the above compound inDCM gives the title compound as a hydrochloride salt as beige powder.

Mp: 236° C. (decomposition).

¹H NMR (DMSO-d₆+MeOD-d₄ (35:1), 300 MHz) δ 8.45 (d, 2H, J=1.5 Hz), 7.71(d, 2H, J=8.7 Hz), 7.59 (dd, 2H, J=8.7, 1.5 Hz), 4.59 (m, 2H), 3.75 (m,1H), 2.91 (m, 4H), 2.80-2.45 (m, 6H).

(±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-aminopropyl]-piperazine-1-carboxylictert-butyl ester is obtained from the treatment of a solution of Example79 (0.05 g, 0.088 mmol) in anhydrous TMOF (1 mL) with ammonia (1.8 mL,0.5M in 1,4-dioxane). The resulting mixture is stir at 60° C. for 3hours then at 10° C. were added MeOH (2 mL) and NaBH₄ (0.12 g). After 15hours of stirring at rt the reaction mixture is quenched with asaturated aqueous solution of sodium hydrogenocarbonate (15 mL),extracted with Et₂O (3×25 mL). After drying over MgSO₄ and evaporationunder reduced pressure the crude compound is purified via flashchromatography on a 2×20 cm² SiO₂ column using EtOAc/MeOH/NH₃ (25%aqueous) (100/10/1.25) as eluant to give(±)-4-[3-(3,6-dibromocarbazol-9-yl)-2-aminopropyl]-piperazine-1-carboxylictert-butyl ester (0.029 g, 0.051 mmol, 58% yield) as a yellow oil.

¹H NMR (CDCl₃, 300 MHz) δ 8.09 (d, 2H, J=1.9 Hz), 7.52 (dd, 2H, J=8.7,1.9 Hz), 7.34 (d, 2H, J=8.7 Hz), 4.25 (dd, 1H, J=14.7, 4.5 Hz), 4.11(dd, 1H, J=14.7, 7.9 Hz), 3.51 (m, 1H), 3.39 (m, 4H), 2.37 (m, 6H), 1.46(br. s, 2H), 1.43 (s, 9H).

Example 116(±)-N-Benzyl-N-[2-(3,6-dibromocarbazol-9-yl)-1-(piperazin-1-ylmethyl)ethyl]amine

The same method as employed in the preparation of Example 3 but startingfrom(±)-4-[3-(3,6-dibromocarbazol-9-yl)-2-benzylaminopropyl]-piperazine-1-carboxylictert-butyl ester gives the title compound in a 85% yield as a colorlessoil. Slow addition of HCl (1M) into a solution of the above compound inDCM gives the title compound as a hydrochloride salt as a beige powder.

Mp.: 218° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.52 (d, 2H, J=1.5 Hz), 7.74 (d, 2H,J=8.7 Hz), 7.66 (dd, 2H, J=8.7, 1.5 Hz), 7.52 (m, 2H), 7.43 (m, 3H),5.00 (dd, 1H, J=15.1, 4.9 Hz), 4.84 (dd, 1H, J=15.1, 8.9 Hz), 4.30 (brs, 2H), 3.79 (m, 1H), 3.02 (dd, 1H, J=13.6, 8.1 Hz), 2.72 (br s, 4H),2.45-2.15 (m, 5H).

(±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-benzylaminopropyl]-piperazine-1-carboxylictert-butyl ester is obtained from the treatment of a solution of Example79 (0.05 g, 0.088 mmol) in anhydrous TMOF (1 mL) with benzylamine (0.100mL) at 60° C. for 3 hours. MS monitoring showed formation of the imineintermediate. Then at rt were added MeOH (1 mL) and NaBH₄ (0.120 g). Theresulting mixture is allowed to stir at rt for 16 hours. The reactionmixture is quenched with a saturated aqueous solution of sodiumhydrogenocarbonate (15 mL). Extraction with Et₂O (3×25 mL), drying overMgSO₄, evaporation under reduced pressure and flash chromatography(SiO₂, 2×16 cm² column), Et₂O/MeOH (100/5)) as eluting solvent gives(±)-4-[3-(3,6-dibromocarbazol-9-yl)-2-benzylamino-propyl]-piperazine-1-carboxylictert-butyl ester as a yellow oil in a 55% yield.

¹H NMR (CDCl₃, 300 MHz) δ 8.12 (d, 2H, J=1.8 Hz), 7.53 (dd, 2H, J=8.9,1.8 Hz), 7.26 (m, 5H), 7.04 (m, 2H), 4.29 (dd, 1H, J=14.7, 6.2 Hz), 4.17(dd, 1H, J=14.7, 6.1 Hz), 3.71 (d, 1H, J=13.6 Hz), 3.52 (d, 1H, J=13.6Hz), 3.31 (m, 4H), 3.19 (m, 1H), 2.42 (dd, 1H, J=12.2, 9.2 Hz), 2.17 (m,5H), 2.04 (br. s, 1H), 1.44 (s, 9H).

Example 117(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(2-morpholin-4-yl-2-oxoethyl)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 73 butstarting from Intermediate 15 and 4-(piperazin-1-ylacetyl)morpholinegives the hydrochloride salt of the title compound as a white powder(0.137 g, 0.21 mmol) in a 81% yield.

Mp: 255° C. (decomposition).

¹H NMR (DMSO-d₆+D₂O, 300 MHz) δ 8.41 (d, 2H, J=1.5 Hz), 7.63 (d, 2H,J=8.7 Hz), 7.58 (dd, 2H, J=8.7, 1.5 Hz), 4.33 (m, 3H), 4.02 (br. s, 2H),3.76 (m, 8H), 3.55 (m, 4H), 3.44 (m, 2H), 3.34 (m, 4H).

Example 118(±)-1-(3,6-Dibromocarbazol-9-yl)-3-{[4-(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-yl)methyl]piperazin-1-yl}propan-2-ol

A solution of Example 5 (0.040 g, 0.029 mmol) in DMF (1 mL) is treatedwith8-bromo-methyl-4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene(BODIPY 493/503 methyl bromide) (0.01 g, 1 equiv.) and DIEA (14.9 μL).The reaction mixture is kept in the dark and is allowed to stir at rtfor 2 d. After evaporation of the solvent the crude compound is purifiedvia flash chromatography using a 2×24 cm² column of SiO₂ andDCM/MeOH/NH₃ (25% aqueous) (90/1/0.1) as eluting solvent to give thetitle compound (Rf=0.24) as a red solid in a 95% yield.

FI-MS (APCI): m/z observed in a negative mode: 726.2.

HPLC (C8 Waters Symmetry Column, 4.6×50 mm², 254 nm, 2 mL/min, ACN/H₂Ogradient with 0.1% TFA): retention time of 6.26 min.

Example 119(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methylacetamide)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 118 butstarting fromN-[(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl]iodoacetamide(BODIPY FL C₁-IA) gives the title compound (Rf=0.45, SiO₂, ACN/NH₃ (25%aqueous) (12/1)) as a red solid in a 84% yield.

FI-MS (APCI): m/z observed in a positive mode: 757.2

FI-MS (APCI): m/z observed in a negative mode: 755.0.

HPLC (C8 Waters Symmetry Column, 4.6×50 mm², 254 nm, 2 mL/min, ACN/H₂Ogradient with 0.1% TFA): retention time of 5.47 min.

Example 120(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(N-(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl)acetamide)piperazin-1-yl]propan-2-ol

The same method as employed in the preparation of Example 118 butstarting fromN-(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl)iodoacetamide(BODIPY 507/545 IA) gives the title compound (Rf=0.32, SiO₂,DCM/MeOH/NH₃ (25% aqueous) (90/7/1)) as a red solid in a 68% yield.

FI-MS (Turbo Scan): m/z observed in a negative mode: 769.4.

HPLC (C8 Waters Symmetry Column, 4.6×50 mm², 254 nm, 2 mL/min, ACN/H₂Ogradient with 0.1% TFA): retention time of 5.64 min.

Example 121(±)-4-[({-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxypropyl]piperazin-1-yl}acetyl)amino]-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoicacid

The same method as employed in the preparation of Example 118 butstarting from2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)-4-[(iodoacetyl)amino]benzoic acidgives after HPLC purification the title compound (Waters Nova-Pack HRC18 column, 25×100 mm², 6 uM, 60 Å, 30 mL/min, ACN/H₂O gradient with0.1% TFA, 254 nm, retention time of 5.1 min) as an orange powder in a90% yield.

M.p.: 107° C.

FI-MS (APCI): m/z observed in a negative mode: 853.2

Example 122(±)-4-({-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxypropyl]piperazin-1-yl}methyl)-6,7-dimethoxy-2H-chromen-2-one

The same method as employed in the preparation of Example 118 butstarting from 4-(bromomethyl)-6,7-dimethoxy-2H-chromen-2-one gives afterHPLC purification the title compound (Waters Nova-Pack HR C18 column,25×100 mm², 6 μM, 60 Å, 30 mL/min, ACN/H₂O gradient with 0.1% TFA, 254nm, retention time of 5.2 min) as a yellow powder in a 32% yield.

M.p.: 112° C.

FI-MS (APCI): m/z observed in a negative mode: 683.8.

FI-MS (APCI): m/z observed in a positive mode: 686.2.

Analysis for C₃₁H₃₃Br₂N₃O₅. 2(C₂F₃O₂):

Calculated: C, 46.02; H, 3.64; N, 4.60;

Found: C, 46.01; H, 3.69; N, 4.68°/:

Example 123(±)-1-(3,6-Dibromocarbazol-9-yl)-3-{4-[3-(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-yl)-propionyl]piperazin-1-yl}propan-2-ol

A solution of Example 5 (0.017 g, 0.036 mmol) in DMF (1 mL) is treatedwith4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-propionicacid, succinimidyl ester (BODIPY 493/503, SE) (0.015 g, 1 equiv.). Theresulting solution is kept in the dark and allowed to stir at rt for 3hours. After evaporation of the solvent the crude compound is purifiedvia preparative HPLC (Waters Nova Pack HRC18, 6 μM, 25×100 mm²,retention time of 5.9 min, using a H₂O/ACN/TFA (0.1%) gradient to givethe title compound as a trifluoroacetate salt as an orange powder in a95% yield.

M.p.: 160° C.

FI-MS (APCI): m/z observed in a positive mode: 770.2.

¹H NMR (DMSO-d₆, 300 MHz) δ 9.67 (br s, 1H), 8.49 (s, 2H), 7.72-7.54 (m,4H), 6.24 (s, 2H), 6.04-5.84 (br s, 1H), 4.5-4.3 (m, 4H), 4.14-3.88 (m,1H), 3.32-3.12 (m, 6H), 3.4-2.83 (m, 3H), 2.82-2.55 (m, 3H), 2.40 (s,6H), 2.36 (s, 6H).

Example 124(±)-1-[4-(4-Nitro-2,1,3-benzoxadiazol-7-yl)-piperazin-1-yl]-3-(3,6-dibromocarbazol-9-yl)-propan-2-ol

A solution of Example 5 (0.060 g, 0.128 mmol) in DMSO (1 mL) is treatedwith 4-fluoro-7-nitro-benz-2-oxa-1,3-diazole (0.024 g, 1 equiv.). Theresulting solution is kept in the dark and allowed to stir at rt for 2hours. After evaporation of the solvent the crude compound is purifiedvia preparative HPLC (Waters Nova Pack HRC18, 6 μM, 25×100 mm²,retention time of 5.52 min, using a H₂O/ACWITA (0.1%) gradient to givethe title compound as a trifluoroacetate salt as an orange powder in a87% yield.

M.p.: 247° C.

FI-MS (Turbo Ion): m/z observed in a positive mode: 631.2.

FI-MS (Turbo Ion): m/z observed in a negative mode: 629.0.

Example 125 Preparation of a Pharmaceutical Formulation

The following formulation examples illustrate representativepharmaceutical compositions of this invention containing carbazolederivatives according to formula I. The present invention, however, isnot limited to the following pharmaceutical compositions.

Formulation 1—Tablets

A compound of formula I is admixed as a dry powder with a dry gelatinbinder in an approximate 1:2 weight ration. A minor amount of magnesiumstearate is added as a lubricant. The mixture is formed into 240-270 mgtablets (80-90 mg of active piperazine derivatives of carbazoleaccording to formula I per tablet) in a tablet press.

Formulation 2—Capsules

A compound of formula I is admixed as a dry powder with a starch diluentin an approximate 1:1 weight ratio. The mixture is filled into 250 mgcapsules (125 mg of active piperazine derivatives of carbazole accordingto formula I per capsule).

Formulation 3—Liquid

A compound of formula I (1250 mg), sucrose (1.75 g) and xanthan gum (4mg) are blended, passed through a No. 10 mesh U.S. sieve, and then mixedwith a previously made solution of microcrystalline cellulose and sodiumcarboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate (10mg), flavor, and color are diluted with water and added with stirring.Sufficient water is then added to produce a total volume of 5 mL.

Formulation 4—Tablets

The compound of formula I is admixed as a dry powder with a dry gelatinbinder in an approximate 1:2 weight ratio. A minor amount of magnesiumstearate is added as a lubricant. The mixture is formed into 450-900 mgtablets (150-300 mg of active piperazine derivatives of carbazoleaccording to formula I) in a tablet press.

Formulation 5—Injection

The compound of formula I is dissolved in a buffered sterile salineinjectable aqueous medium to a concentration of approximately 5 mg/ml.

In the following the present invention shall be illustrated by means ofsome examples which are not construed to be viewed as limiting the scopeof the invention.

Example 126 Biological Assays

a) Production of Recombinant Bax

Human Bax-α lacking 20 amino acids at the COOH-terminus is expressed asa GST fusion protein or a His-tagged protein in Escherichia coli, andthe protein is purified from the soluble cell fraction. In brief, theGST-Bax fusion protein is applied to a glutathione-Sepharose column, andBax is released by cleavage with thrombin (0.6 U/mL). Bax issubsequently purified on heparin-Sepharose, followed by fast proteinliquid chromatography (FPLC) Mono Q. His-tagged Bax is purified on aNi-nitriloacetic acid-agarose column followed by FPLC MonoQ:

b) Isolation of Mitochondria

Mitochondria are isolated from mouse liver cells by differentialcentrifugation. Cells are broken with a dounce homogenizer and thesuspension is centrifuged at 2,000 g in an Eppendorf centrifuge at 4° C.This procedure is repeated until almost all the cells are broken.Supernatants from each step are pooled before centrifugation at 13,000 gat 4° C. for 10 min. The pellet is resuspended in 40 mL MB buffer andcentrifuged at 2000 g for 2 min. The supernatant is removed andcentrifuged at 13 kg for 4 min. The mitochondria are recovered in the 13k pellet and resuspended in MB buffer at a density of 30 OD600 nm/mL.

c) In Vitro Assay for Cytochrome c Release (Mitochondria Cytochrome cRelease Triggered by Bax Activation)

Mitochondria (30 μg) from mouse liver are incubated with 200 nMrecombinant Bax in the presence of various compounds (5 μM) in 200 μL ofKCl buffer for 20 min at 30° C. and are then centrifuged for 4 min at13,000 g at 4° C. Mitochondrial pellets corresponding to 1.5 μg proteinsare separated by SDS-PAGE using 4-20% Tris-Gly gels (NOVEX) and theirrespective contents of cytochrome c are estimated by Western blottingusing polyclonal anti-cytochrome c antibody (dilution 1:2,500).Antigen-antibody complexes are detected using horseradishperoxidase-conjugated goat anti-rabbit IgG and enhance chemiluminescencedetection reagents. The cytochrome c bands are scanned and quantifiedusing a Bio-Rad (GS-700 Imaging Densitometer).

d) Effect of Compounds according to formula I onto the Release ofCytochrome c Triggered by Bid-Induced Bax Activation (MitochondriaCytochrome c Release Triggered by Bid-Induced Bax Activation)

Concerning the Bid-induced activation of Bax leading to mitochondrialCytochrome C release, it is referred to the description of Martinou etal. in The Journal of Cell Biology, Vol. 144, No. 5, Mar. 8, 1999, pages891-901. Mitochondria isolated from HeLa cells are incubated for 15 minat 30° C. in 100 μA of KCl buffer in the presence or absence of 10 nMrecombinant Bid. The various compounds (10 μM) are pre-incubated for 5min prior to addition of Bid. Following incubation, mitochondria werecentrifuged for 5 min at 13000 g at 4° C. and the supernatant iscollected for cytochrome c analysis. Cytochrome c is detected by Westernblotting. The cytochrome c bands are scanned and quantified using aBio-Rad (GS-700 Imaging Densitometer).

The above set out 2 in vitro assays c) and d) involving thedetermination of mitochondrial cytochrome c release are based onimmunochemical methods using the Western blot analysis. Alternatively,said quantitative cytochrome c determinations could be performed byusing spectrophotometric means:

-   I. by recording the difference between reduced and oxidised    cytochrome c by dual wavelength double beam spectrophotometry;-   by measuring the rather intensive γ or Soret peak in the spectrum of    cytochrome c (ε=100 mM⁻¹ cm⁻¹) is used for rapid and quantitative    determination of the release of cytochrome c from isolated    mitochondria. This technique allows a highly convenient, fast and    reliable quantitative determination of the release of cytochrome c.    e) Sympathetic Neuron Culture and Survival Assay (Neuronal Survival)

Sympathetic neurons from superior cervical ganglia (SCG) of newborn rats(p4) are dissociated in dispase, plated at a density of 104 cells/cm² in48 well MTT plates coated with rat tail collagen, and cultured inLeibowitz medium containing 5% rat serum, 0.75 g/ml NGF 7S (BoehringerMannheim Corp., Indianapolis, Ind.) and arabinosine 105M. Cell death isinduced at day 4 after plating by exposing the culture to mediumcontaining 10 g/ml of anti NGF antibody (Boehringer Mannheim Corp.,Indianapolis, Ind.) and no NGF or arabinosine, in the presence orabsence of piperazine derivatives of carbazole inhibitors according toformula I. 24 hours after cell death induction, determination of cellviability is performed by incubation of the culture for 1 hour, at 37°C. in 0.5 mg/ml of 3-(4,5-dimethyl-thiazol-2-yl)2,5diphenyl tetrazoliumbromide (MTT). After incubation in MTT, cells are re-suspended in DMSO,transferred to a 96 MTT plate and cell viability is evaluated bymeasuring optical density at 590 nm.

f) Biological Results—Discussion

The activities of the piperazine derivatives of carbazoles claimed inthe formula I were assessed using the above described in vitrobiological assays. Representative values are given in the table shownbelow:

Mitochondria Cytochrome Mitochondria c Release Cytochrome c Triggered byRelease Bid-Induced Triggered Bax Acti- by Bax Neuronal vationActivation Survival^((a)) Compound (% Inhibition)^((a)) (%Inhibition)^((b)) (%) 23 68 51 45 27 88 40 40 32 98 43 30 45 37n.a.^((c)) 60 60 60 n.a.^((c)) 40 62 99 n.a.^((c)) 30 73 74 n.a.^((c))30 80 73 n.a.^((c)) 20 110 87 n.a.^((c)) 30 ^((a))Compounds were testedat 10 μM ^((b))Compounds were tested at 5 μM ^((c))n.a. = not available

The test compounds are among those described in the Examples 1-124 andtheir designation is derived therefrom. The above indicated values incolumn 2 and 3 refer to the inhibition (in %) of the mitochondrialcytochrome c release upon using the corresponding test compounds.

From the above table, it is derived that the test compounds according tothe formula I do have a significant effect on the inhibition of releaseof cytochrome c. According to a preferred embodiment the testedcompounds of formula I display an inhibition of the cytochrome c releaseof at least 40%, more preferred of at least 60% when tested at aconcentration of between 2-50 μM, preferably between 5-20 μM and mostpreferred at 5-10 μM.

According to a preferred embodiment, the tested compounds display aneuronal survival rate of at least 30%, preferably of at least 40%.

1. A compound according to formula I

as well as a pharmaceutically acceptable salt thereof, in racemic formor in an enantiomeric excess, wherein R⁰ and R¹ are selectedindependently from each other and are cyano, sulfonyl of the formula—SO₂—R where R is H, aryl, heteraryl, C₁-C₆ alkyl or C₁-C₆ alkylsubstituted with a halogen, sulfoxy, substituted or unsubstitutedC₁-C₆-thioalkoxy, nitro, primary, secondary or tertiary amine orsulfonamide, aminocarbonyl, amino-thiocarbonyl, hydroxy, substituted orunsubstituted C₁-C₆-alkoxy, aryloxy, heteroaryloxy, carboxylic amide,alkoxycarbonyl, carboxylic ester, carboxylic acid, substituted orunsubstituted C₁-C₆-alkyl carbonyl, substituted or unsubstitutedarylcarbonyl or heteroarylcarbonyl, substituted or unsubstitutedsaturated or unsaturated C₃-C₈-cycloalkylcarbonyl, substituted orunsubstituted C₁-C₆-alkyl, substituted or unsubstituted C₂-C₆-alkenyl,substituted or unsubstituted C₂-C₆-alkynyl, substituted or unsubstitutedaryl or heteroaryl, substituted or unsubstituted 3-8 membered saturatedor unsaturated cyclic alkyl, a lipophilic substituent selected from thegroup consisting of bromine, aryl, and C₁-C₆-alkyl, or a fluorescentmoiety selected from the group consisting of(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl)acetamide,[(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl]acetamide,(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacen-8-yl)methyl,2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)-4-(aminoacetyl)benzoic acid,(6,7-dimethoxy-2H-chromen-2-one)-4-methyl,4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-propionyl,and 4-nitro-(2,1,3-benzoxadiazol)-7-yl; R² is C₁-C₆-alkylaryl,C₁-C₆-alkylheteroaryl, an aryl, or a 5-6-membered heterocyclic groupcontaining at least one of oxygen, nitrogen and sulfur, wherein the aryland heterocyclic group are substituted by at least one C(═O)ORa,trifluoromethyl, trifluoromethoxy, ORa, OC(═O)Ra, OC(═O)Rc, NRaRb,CH₂—NRaRb, NO₂, cyano, halogen, SO₂NRaRb, SO₂NRaRc, NRaSO₂Ra, NRaSO₂Rc,C₁-C₆-alkyleneC(═O)ORa, and OSO₂-trifluoromethyl, or R² isC₁-C₆-alkylaryl or C₁-C₆-alkylheteroaryl containing at least oneheteroatom selected from oxygen, nitrogen and sulfur, wherein saidC₁-C₆-alkylaryl or C₁-C₆-alkylheteroaryl is substituted by at least oneC(═O)ORa, trifluoromethyl, trifluoromethoxy, ORa, ORc, SRa, SRc, OC(═O)Ra, OC(═O) Rc, NRaRb, CH₂—NRaRb, NO₂, cyano, halogen, SO₂NRaRb,SO₂NRaRc, NRaSO₂Ra, NRaSO₂Rc, C₁-C₆-alkyleneC(═O)ORa orOSO₂-trifluoromethyl; Ra and Rb, in R², are independently hydrogen orC₁-C₆-alkyl, optionally substituted by at least one halogen, aC₁-C₆-alkoxy or an amino group and Rc, in R², is an unsubstituted orsubstituted phenyl, an unsubstituted or substituted benzyl or a3-8-membered unsubstituted or substituted saturated 3-8-membered cyclicalkyl; R³ is hydrogen, halogen, substituted or unsubstitutedC₁-C₆-alkyl, substituted or unsubstituted C₂-C₆-alkenyl, substituted orunsubstituted C₂-C₆-alkynyl, substituted or unsubstituted aryl orhetero-aryl, substituted or unsubstituted 3-8 membered saturated andunsaturated cyclic alkyl, alkoxycarbonyl, carboxylic amide,C₁-C₆-alkoxy, substituted or unsubstituted aryloxy, substituted orunsubstituted hetero-aryloxy, hydroxy, substituted or unsubstitutedC₁-C₆-alkyl carbonyl, substituted or unsubstituted arylcarbonyl orheteroarylcarbonyl, substituted or unsubstituted saturated orunsaturated C₄-C₈-cycloalkylcarbonyl, or an oxo (═O) group; k and l are1; X is a group of the formula —(CR′R″)—, wherein R′ and R″ arehydrogen, hydroxyl or a substituted or unsubstituted C₁-C₆ alkoxy,whereby at least one of R′ and/or R″ is not hydrogen but hydroxyl or asubstituted or unsubstituted C₁-C₆-alkoxy; m and n are independentlyfrom each other an integer from 1 to 3 and o is an integer from 0 to 8.2. The compound according to claim 1, wherein R⁰ and R¹ of formula I areselected independently from each other and are cyano, substituted orunsubstituted and C₁-C₆-alkyl, substituted or unsubstituted 3-8 memberedsaturated or unsaturated cyclic alkyl, C(═O)ORa, C(═O)NRaRb, C(═O)NRaRc,C(═O)Ra, C(═O)Rc, CRa(═N—N-Rb), CRa(═N—N-Rc), CRa(═N—O-Rb),trifluoromethyl, trifluoromethoxy, ORa, ORc, NRaRb, NRaRc,NRaC(═O)NRaRb, NRaC(═O)NRaRc, NRaC(═O)Rb, NRaC(═O)Rc, OC(═O)Ra,OC(═O)Rc, NRa(SO₂Rb), NRa(SO₂Rc), SO₂NRaRb, SO₂NRaRc, NO₂, CH₂NRaRb,CH₂NRaRc, CH₂NRaC(═O)NRaRb, SRa, SRc, CH₂NRaC(═O)NRaRc, CH₂NRaC(═O)Rb,CH₂NRaC(═O)Rc, CH₂NRa(SO₂Rb), CH₂NRa(SO₂Rc), OSO₂-trifluoromethyl; oraryl or a 5-6-membered heteroaryl or heterocyclic group wherein at leastone heteroatom in the 5-6 membered heteroaryl or heterocyclic group isoxygen, nitrogen or sulfur, said aryl or heteroaryl or heterocyclicgroup being optionally substituted by at least one C₁-C₆-alkyl,C(═O)ORa, trifluoromethyl, trifluoromethoxy, ORa, ORc, SRa, SRc,OC(═O)Ra, OC(═O)Rc, NRaRb, CH₂—NRaRb, NO₂, cyano, halogen, SO₂NRaRb,SO₂NRaRc, NRaSO₂Ra, NRaSO₂Rc, C₁-C₆-alkyleneC(═O)ORa,OSO₂-trifluoromethyl; with Ra and Rb, in R¹, R⁰ or both, being the sameor different, independently selected from hydrogen and C₁-C₆-alkyl,being optionally substituted by at least one halogen, C₁-C₆-alkoxy oramino group; and Rc, in R¹, R⁰ or both, is an unsubstituted orsubstituted phenyl, an unsubstituted or substituted benzyl, a3-8-membered unsubstituted or substituted saturated 3-8-membered cyclicalkyl, a lipophilic substituents selected from the group consisting ofbromine, aryl, and C₁-C₆-alkyl, or a fluorescent moiety selected fromthe group consisting of(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl)acetamide,[(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl]acetamide,(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacen-8-yl)methyl,2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)-4-(aminoacetyl)benzoic acid,(6,7-dimethoxy-2H-chromen-2-one)-4-methyl,4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-propionyl,and 4-nitro-(2,1,3-benzoxadiazol)-7-yl.
 3. The compound according toclaim 1, wherein R² of formula I is C₁-C₆-alkylaryl,C₁-C₆-alkylheteroaryl, or an aryl, wherein the aryl group is substitutedby at least one of C(═O)ORa, trifluoromethyl, trifluoromethoxy, ORa,OC(═O)Ra, OC(═O)Rc, NRaRb, CH₂—NRaRb, NO₂, cyano, halogen, SO₂NRaRb,SO₂NRaRc, NRaSO₂Ra, NRaSO₂Rc, C₁-C₆-alkyleneC(═O)ORa, andOSO₂trifluoromethyl; Ra and Rb, in R², are independently hydrogen orC₁-C₆-alkyl, optionally substituted by at least one halogen, aC₁-C₆-alkoxy or an amino group and Rc, in R², is an unsubstituted orsubstituted phenyl, an unsubstituted or substituted benzyl or a3-8-membered unsubstituted or substituted saturated 3-8-membered cyclicalkyl.
 4. The compound according to claim 1, wherein R³ of formula I ishydrogen, C₁-C₆-alkyl, ORa, ORc, C(═O)ORa, C(═O)ORc C(═O)NRaRb,C(═O)NRaRc, C(═O)Ra, C(═O)Rc, RaC(═O)NRaRc, RaC(═O)Rb, RaC(═O)Rc,Ra(SO₂Rb), Ra(SO₂Rc), (═O); or R³ of formula I is an aryl or a5-6-membered heterocyclic group wherein at least one heteroatom in the5-6 membered heterocyclic group is selected from the group consisting ofoxygen, nitrogen and sulfur, both the aryl, heterocyclic group beingoptionally substituted by at least one C₁-C₆-alkyl, C(═O)ORa, C(═O)ORc,trifluoromethyl, trifluoromethoxy, ORa, OC(═O)Ra, OC(═O)Rc, NRaRb,CH₂—NRaRb, NO₂, cyano, halogen, SO₂NRaRb, SO₂NRaRc, NRaSO₂Ra, NRaSO₂Rc,C₁-C₆-alkyleneC(═O)ORa, OSO₂trifluoromethyl; whereby Ra and Rb are thesame or different and they are independently selected from hydrogen andC₁-C₆-alkyl, being optionally substituted by at least one halogen, aC₁-C₆-alkoxy or an amino group; and whereby, Rc does represent thereinan unsubstituted or substituted phenyl, an unsubstituted or substitutedbenzyl or a 3-8-membered unsubstituted or substituted saturated3-8-membered cyclic alkyl.
 5. The compound according to claim 1, whereinR⁰ and R¹ represent a lipophilic substitutent selected from the groupconsisting of bromine, aryl, and C₁-C₆-alkyl.
 6. The compound accordingto claim 1, wherein R³ is hydrogen or C₁-C₆-alkyl.
 7. The compoundaccording to claim 1, wherein R² is(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl)acetamide,[(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl]acetamide,(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacen-8-yl)methyl,2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)-4-(aminoacetyl)benzoic acid,(6,7-dimethoxy-2H-chromen-2-one)-4-methyl,4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-propionyl,or 4-nitro-(2,1,3-benzoxadiazol)-7-yl.
 8. The compound according toclaim 1, which is in an enantiomeric excess of at least 52% ee.
 9. Thecompound according to claim 1 selected from the following group:(±)-1-[(4-Fluorophenyl)-piperazin-1-yl]-3-(3,6-dibromocarbazol-9-yl)-propan-2-ol;(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(thien-3-ylmethyl)piperazin-1-yl]propan-2-ol;(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-methoxybenzyl)piperazin-1-yl]propan-2-ol;(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(4-tert-butylbenzyl)piperazin-1-yl]propan-2-ol;(±)-4-[3-(3,6-Dibromocarbazol-9-yl)-2-hydroxypropyl]-N-(4-fluorophenyl)piperazine-1-carboxamide;(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(3,4-dichlorobenzyl)piperazin-1-yl]propan-2-ol;(±)-1-(3,6-Dibromo-carbazol-9-yl)-3-[4-(2-morpholin-4-ethyl)piperazin-1-yl]propan-2-ol;(±)-1-(3,6-Dibromocarbazol-9-yl)-3-[4-(3-piperidin-1-yl-propyl)piperazin-1-yl]propan-2-ol;(±)-1-(3,6-dibromo-carbazol-9-yl)-3-[4-(4-fluoro-benzyl)-piperazin-1yl]-propan-2-ol;(±)-1-(3,6-dibromo-carbazol-9-yl)-3-[4-(3-phenyl-propyl)-piperazin-1-yl]-propan-2-ol;(±)-1-(3,6-dibromo-carbazol-9-yl)-3-[4-(thien-2-ylmethyl)piperazin-1-yl]propan-2-ol;(±)-1-(3,6-dibromocarbozyl-9-yl)-3-[4-(pyridine-3-ylmethyl)piperazin-1-yl]propan-2-ol;(±)-1-(3,6-dibromocarbazol-9-yl)-3-[4-trifluoromethylbenzyl]piperazin-1-yl]propan-2-ol;and(±)-1-(3,6-Dibromocarbazol-9-yl)-3-{[4-(4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-yl)methyl]piperazin-1-yl}propan-2-ol.10. A pharmaceutical composition comprising at least one compoundaccording to claim 1 and a pharmaceutically acceptable carrier, diluentor excipient thereof.
 11. The compound according to claim 1, which is inan enantiomeric excess of at least 92-98% ee.
 12. A pharmaceuticalcomposition comprising at least one compound according to claim 2 and apharmaceutically acceptable carrier, diluent or excipient thereof.
 13. Apharmaceutical composition comprising at least one compound according toclaim 3 and a pharmaceutically acceptable carrier, diluent or excipientthereof.
 14. A pharmaceutical composition comprising at least onecompound according to claim 4 and a pharmaceutically acceptable carrier,diluent or excipient thereof.
 15. A pharmaceutical compositioncomprising at least one compound according to claim 5 and apharmaceutically acceptable carrier, diluent or excipient thereof.
 16. Apharmaceutical composition comprising at least one compound according toclaim 6 and a pharmaceutically acceptable carrier, diluent or excipientthereof.
 17. A pharmaceutical composition comprising at least onecompound according to claim 7 and a pharmaceutically acceptable carrier,diluent or excipient thereof.
 18. A pharmaceutical compositioncomprising at least one compound according to claim 8 and apharmaceutically acceptable carrier, diluent or excipient thereof.
 19. Apharmaceutical composition comprising at least one compound according toclaim 9 and a pharmaceutically acceptable carrier, diluent or excipientthereof.
 20. A pharmaceutical composition comprising at least onecompound according to claim 11 and a pharmaceutically acceptablecarrier, diluent or excipient thereof.